Ketoamides with cyclic P4&#39;S as inhibitors of NS3 protease of hepatitis C virus

ABSTRACT

The present invention discloses novel compounds which have HCV protease inhibitory activity as well as methods for preparing such compounds. In another embodiment, the invention discloses pharmaceutical compositions comprising such compounds as well as methods of using them to treat disorders associated with the HCV protease.

This application claims priority from U.S. provisional patentapplication Ser. No. 60/548,506 filed Feb. 27, 2004.

FIELD OF THE INVENTION

The present invention relates to novel hepatitis C virus (“HCV”)protease inhibitors, pharmaceutical compositions containing one or moresuch inhibitors, methods of preparing such inhibitors and methods ofusing such inhibitors to treat hepatitis C and related disorders. Thisinvention additionally discloses novel macrocyclic compounds asinhibitors of the HCV NS3/NS4a serine protease.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) is a (+)-sense single-stranded RNA virus thathas been implicated as the major causative agent in non-A, non-Bhepatitis (NANBH), particularly in blood-associated NANBH (BB-NANBH)(see, International Patent Application Publication No. WO 89/04669 andEuropean Patent Application Publication No. EP 381 216). NANBH is to bedistinguished from other types of viral-induced liver disease, such ashepatitis A virus (HAV), hepatitis B virus (HBV), delta hepatitis virus(HDV), cytomegalovirus (CMV) and Epstein-Barr virus (EBV), as well asfrom other forms of liver disease such as alcoholism and primary biliarcirrhosis.

Recently, an HCV protease necessary for polypeptide processing and viralreplication has been identified, cloned and expressed. (See, e.g., U.S.Pat. No. 5,712,145). This approximately 3000 amino acid polyproteincontains, from the amino terminus to the carboxy terminus, anucleocapsid protein (C), envelope proteins (E1 and E2) and severalnon-structural proteins (NS1, 2, 3, 4a, 5a and 5b). NS3 is anapproximately 68 kda protein, encoded by approximately 1893 nucleotidesof the HCV genome, and has two distinct domains: (a) a serine proteasedomain consisting of approximately 200 of the N-terminal amino acids;and (b) an RNA-dependent ATPase domain at the C-terminus of the protein.The NS3 protease is considered a member of the chymotrypsin familybecause of similarities in protein sequence, overall three-dimensionalstructure and mechanism of catalysis. Other chymotrypsin-like enzymesare elastase, factor Xa, thrombin, trypsin, plasmin, urokinase, tPA andPSA. The HCV NS3 serine protease is responsible for proteolysis of thepolypeptide (polyprotein) at the NS3/NS4a, NS4a/NS4b, NS4b/NS5a andNS5a/NS5b junctions and is thus responsible for generating four viralproteins during viral replication. This has made the HCV NS3 serineprotease an attractive target for antiviral chemotherapy. The inventivecompounds can inhibit such protease. They also can modulate theprocessing of hepatitis C virus (HCV) polypeptide.

It has been determined that the NS4a protein, an approximately 6 kdapolypeptide, is a co-factor for the serine protease activity of NS3.Autocleavage of the NS3/NS4a junction by the NS3/NS4a serine proteaseoccurs intramolecularly (i.e., cis) while the other cleavage sites areprocessed intermolecularly (i.e., trans).

Analysis of the natural cleavage sites for HCV protease revealed thepresence of cysteine at P1 and serine at P1′and that these residues arestrictly conserved in the NS4a/NS4b, NS4b/NS5a and NS5a/NS5b junctions.The NS3/NS4a junction contains a threonine at P1 and a serine at P1′.The Cys→Thr substitution at NS3/NS4a is postulated to account for therequirement of cis rather than trans processing at this junction. See,e.g., Pizzi et al. (1994) Proc. Natl. Acad. Sci. (USA) 91:888–892,Failla et al. (1996) Folding & Design 1:35–42. The NS3/NS4a cleavagesite is also more tolerant of mutagenesis than the other sites. See,e.g., Kollykhalov et al. (1994) J. Virol. 68:7525–7533. It has also beenfound that acidic residues in the region upstream of the cleavage siteare required for efficient cleavage. See, e.g., Komoda et al. (1994) J.Virol. 68:7351–7357.

Inhibitors of HCV protease that have been reported include antioxidants(see, International Patent Application Publication No. WO 98/14181),certain peptides and peptide analogs (see, International PatentApplication Publication No. WO 98/17679, Landro et al. (1997) Biochem.36:9340–9348, Ingallinella et al. (1998) Biochem. 37:8906–8914,Llinàs-Brunet et al. (1998) Bioorg. Med. Chem. Lett. 8:1713–1718),inhibitors based on the 70-amino acid polypeptide eglin c (Martin et al.(1998) Biochem. 37:11459–11468, inhibitors affinity selected from humanpancreatic secretory trypsin inhibitor (hPSTI-C3) and minibodyrepertoires (MBip) (Dimasi et al. (1997) J. Virol. 71:7461–7469),cV_(H)E2 (a “camelized” variable domain antibody fragment) (Martin etal. (1997) Protein Eng. 10:607–614), and α1-antichymotrypsin (ACT)(Elzouki et al.) (1997) J. Hepat. 27:42–28). A ribozyme designed toselectively destroy hepatitis C virus RNA has recently been disclosed(see, BioWorld Today 9(217): 4 (Nov. 10, 1998)).

Reference is also made to the PCT Publications, No. WO 98/17679,published Apr. 30, 1998 (Vertex Pharmaceuticals Incorporated); WO98/22496, published May 28, 1998 (F. Hoffmann-La Roche AG); and WO99/07734, published Feb. 18, 1999 (Boehringer Ingelheim Canada Ltd.).

HCV has been implicated in cirrhosis of the liver and in induction ofhepatocellular carcinoma. The prognosis for patients suffering from HCVinfection is currently poor. HCV infection is more difficult to treatthan other forms of hepatitis due to the lack of immunity or remissionassociated with HCV infection. Current data indicates a less than 50%survival rate at four years post cirrhosis diagnosis. Patients diagnosedwith localized resectable hepatocellular carcinoma have a five-yearsurvival rate of 10–30%, whereas those with localized unresectablehepatocellular carcinoma have a five-year survival rate of less than 1%.

Reference is made to WO 00/59929 (U.S. Pat. No. 6,608,027, Assignee:Boehringer Ingelheim (Canada) Ltd.; Published Oct. 12, 2000) whichdiscloses peptide derivatives of the formula:

Reference is made to A. Marchetti et al, Synlett, S1, 1000–1002 (1999)describing the synthesis of bicylic analogs of an inhibitor of HCV NS3protease. A compound disclosed therein has the formula:

Reference is also made to W. Han et al, Bioorganic & Medicinal Chem.Lett, (2000) 10, 711–713, which describes the preparation of certainα-ketoamides, α-ketoesters and α-diketones containing allyl and ethylfunctionalities.

Reference is also made to WO 00/09558 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to WO 00/09543 (Assignee: Boehringer IngelheimLimited; Published Feb. 24, 2000) which discloses peptide derivatives ofthe formula:

where the various elements are defined therein. An illustrative compoundof that series is:

Reference is also made to U.S. Pat. No. 6,608,027 (Boehringer Ingelheim,Canada) which discloses NS3 protease inhibitors of the type:

wherein the various are defined therein.

Current therapies for hepatitis C include interferon-α (INF_(α)) andcombination therapy with ribavirin and interferon. See, e.g., Beremgueret al. (1998) Proc. Assoc. Am. Physicians 110(2):98–112. These therapiessuffer from a low sustained response rate and frequent side effects.See, e.g., Hoofnagle et al. (1997) N. Engl. J. Med. 336:347. Currently,no vaccine is available for HCV infection.

Reference is further made to WO 01/74768 (Assignee: VertexPharmaceuticals Inc) published Oct. 11, 2001, which discloses certaincompounds of the following general formula (R is defined therein) asNS3-serine protease inhibitors of Hepatitis C virus:

A specific compound disclosed in the afore-mentioned WO 01/74768 has thefollowing formula:

PCT Publications WO 01/77113; WO 01/081325; WO 02/08198; WO 02/08256; WO02/08187; WO 02/08244; WO 02/48172; WO 02/08251; and pending U.S. patentapplication Ser. No. 10/052,386, filed Jan. 18, 2002, disclose varioustypes of peptides and/or other compounds as NS-3 serine proteaseinhibitors of hepatitis C virus. The disclosures of those applicationsare incorporated herein by reference thereto.

There is a need for new treatments and therapies for HCV infection.There is a need for compounds useful in the treatment or prevention oramelioration of one or more symptoms of hepatitis C.

There is a need for methods of treatment or prevention or ameliorationof one or more symptoms of hepatitis C.

There is a need for methods for modulating the activity of serineproteases, particularly the HCV NS3/NS4a serine protease, using thecompounds provided herein.

There is a need for methods of modulating the processing of the HCVpolypeptide using the compounds provided herein.

SUMMARY OF THE INVENTION

In its many embodiments, the present invention provides a novel class ofinhibitors of the HCV protease, pharmaceutical compositions containingone or more of the compounds, methods of preparing pharmaceuticalformulations comprising one or more such compounds, and methods oftreatment or prevention of HCV or amelioration of one or more of thesymptoms of hepatitis C using one or more such compounds or one or moresuch formulations. Also provided are methods of modulating theinteraction of an HCV polypeptide with HCV protease. Among the compoundsprovided herein, compounds that inhibit HCV NS3/NS4a serine proteaseactivity are preferred. The present invention discloses compounds havingthe general structure shown in structural Formula 1:

wherein:

R¹ is H, OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can be the sameor different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroalkyl-,heteroaryl-, cycloalkyl-, heterocyclyl-, arylalkyl-, andheteroarylalkyl;

A and M can be the same or different, each being independently selectedfrom R, OR, NHR, NRR′, SR, SO₂R, and halo; or A and M are connected toeach other (in other words, A-E-L-M taken together) such that themoiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl;

E is C(H) or C(R);

L is C(H), C(R), CH₂C(R), or C(R)CH₂;

R, R′, R², and R³ can be the same or different, each being independentlyselected from the group consisting of H, alkyl-, alkenyl-, alkynyl-,cycloalkyl-, heteroalkyl-, heterocyclyl-, aryl-, heteroaryl-,(cycloalkyl)alkyl-, (heterocyclyl)alkyl-, aryl-alkyl-, andheteroaryl-alkyl-; or alternately R and R′in NRR′are connected to eachother such that NRR′ forms a four to eight-membered heterocyclyl;

and Y is selected from the following moieties:

wherein G is NH or O, and R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹ and R²⁰ can be thesame or different, each being independently selected from the groupconsisting of H, C₁–C₁₀ alkyl, C₁–C₁₀ heteroalkyl, C₂–C₁₀ alkenyl,C₂–C₁₀ heteroalkenyl, C₂–C₁₀ alkynyl, C₂–C₁₀ heteroalkynyl, C₃–C₈cycloalkyl, C₃–C₈ heterocyclyl, aryl, heteroaryl, or alternately: (i)either R¹⁵ and R¹⁶ can be connected to each other to form a four toeight-membered cycloalkyl or heterocyclyl, or R¹⁵ and R¹⁹ are connectedto each other to form a five to eight-membered cycloalkyl orheterocyclyl, or R¹⁵ and R²⁰ are connected to each other to form a fiveto eight-membered cycloalkyl or heterocyclyl, and (ii) likewise,independently, R¹⁷ and R¹⁸ are connected to each other to form a threeto eight-membered cycloalkyl or heterocyclyl,

wherein each of said alkyl, aryl, heteroaryl, cycloalkyl or heterocyclylcan be unsubstituted or optionally independently substituted with one ormore moieties selected from the group consisting of: hydroxy, alkoxy,aryloxy, thio, alkylthio, arylthio, amino, amido, alkylamino, arylamino,alkylsulfonyl, arylsulfonyl, sulfonamido, alkylsulfonamido,arylsulfonamido, keto, carboxy, carbalkoxy, carboxamido,alkoxycarbonylamino, alkoxycarbonyloxy, alkylureido, arylureido, halo,cyano, and nitro.

The above-noted statement “A and M are connected to each other such thatthe moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, a four to eight-membered heterocyclyl, a sixto ten-membered aryl, or a five to ten-membered heteroaryl” can beillustrated in a non-limiting matter as follows. Thus, for example, inthe case where A and M are connected such that the moiety:

shown above in Formula I forms a six-membered cycloalkyl(cyclohexyl),Formula I can be depicted as:

One with ordinary skill in the art will appreciate that similardepictions for Formula I can be arrived at when A and M shown above inthe moiety:

(i.e., M-L-E-A taken together) are connected to form a three, four,seven or eight-membered cycloalkyl, a four to eight-memberedheterocyclyl, a six to ten-membered aryl, or a five to ten-memberedheteroaryl.

In the above-noted definitions of R, R′, R², and R³ preferred alkyl ismade of one to ten carbon atoms, preferred alkenyl or alkynyl is made oftwo to ten carbon atoms, preferred cycloalkyl is made of three to eightcarbon atoms, and preferred heteroalkyl, heteroaryl or heterocycloalkylhas one to six oxygen, nitrogen, sulfur, or phosphorus atoms.

The compounds represented by Formula I, by themselves or in combinationwith one or more other suitable agents disclosed herein, can be usefulfor treating diseases such as, for example, HCV, HIV, AIDS (AcquiredImmune Deficiency Syndrome), and related disorders, as well as formodulating the activity of hepatitis C virus (HCV) protease, preventingHCV, or ameliorating one or more symptoms of hepatitis C. Suchmodulation, treatment, prevention or amelioration can be done with theinventive compounds as well as with pharmaceutical compositions orformulations comprising such compounds. Without being limited to theory,it is believed that the HCV protease may be the NS3 or NS4a protease.The inventive compounds can inhibit such protease. They can alsomodulate the processing of hepatitis C virus (HCV) polypeptide.

DETAILED DESCRIPTION

In an embodiment, the present invention discloses compounds which arerepresented by structural Formula 1 or a pharmaceutically acceptablesalt, solvate or ester thereof, wherein the various moieties are asdefined above.

In another embodiment, R¹ is NR⁹R¹⁰, and R⁹ is H, R¹⁰ is H, alkyl, aryl,heteroalkyl, heteroaryl, cycloalkyl, alkyl-aryl, alkyl-heteroaryl,aryl-alkyl, alkenyl, alkynyl or heteroaryl-alkyl.

In another embodiment, R¹⁰ is selected from the group consisting of:

In another embodiment, R² is selected from the group consisting of thefollowing moieties:

In another embodiment, R³ is selected from the group consisting of:

wherein R³¹ is OH or O-alkyl; and

-   -   R³² is H, C(O)CH₃, C(O)OtBu or C(O)N(H)tBu.        In an additional embodiment, R³ is selected from the group        consisting of the following moieties:

In another embodiment, Y is selected from the group consisting of:

wherein Y³⁰ and Y³¹ are selected from the group consisting of:

Y³² is selected from the group consisting of:

and Y¹² is selected from H, COOH, COOMe, CONH₂, OMe, OH, OCF₃,OCH(CH₃)₂, OC(CH₃)₃, F, Cl, Br, NH₂, NHSO₂CH₃, NHC(O)CH₃, NHCO₂CH₃, NO₂,SO₂NH₂, CF₃, Me, Et, isopropyl, cyclopropyl, t-butyl, phenyl.

In another embodiment, the moiety:

is selected from the following structures:

In an additional embodiment, the moiety:

is selected from the following structures:

In a still additional embodiment, the moiety:

is selected from the following structures:

In a further additional embodiment, R¹ is NHR¹⁰, where R¹⁰ is selectedfrom the group consisting of:

R² is selected from the group consisting of the following moieties:

R³ is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

wherein Y³⁰ and Y³¹ can be the same or different, each beingindependently selected from the group consisting of:

wherein Y³² is selected from the group consisting of:

and Y¹² is selected from H, COOH, COOMe, CONH₂, OMe, OH, OCF₃,OCH(CH₃)₂, OC(CH₃)₃, F, Cl, Br, NH₂, NHSO₂CH₃, NHC(O)CH₃, NHCO₂CH₃, NO₂,SO₂NH₂, CF₃, Me, Et, isopropyl, cyclopropyl, t-butyl, or phenyl; and themoiety:

Representative compounds of the invention which exhibit excellent HCVprotease inhibitory activity are listed later in this Description inTables 1 and 2 along with their biological activity in HCV continuousassay (ranges of Ki* values in nanomolar, nM).

In an additional embodiment, this invention discloses the followingcompounds in Table 3:

TABLE 3

As used above, and throughout this disclosure, the following terms,unless otherwise indicated, shall be understood to have the followingmeanings:

“Patient” includes both human and animals.

“Mammal” means humans and other mammalian animals.

“Alkyl” means an aliphatic hydrocarbon group which may be straight orbranched and comprising about 1 to about 20 carbon atoms in the chain.Preferred alkyl groups contain about 1 to about 12 carbon atoms in thechain. More preferred alkyl groups contain about 1 to about 6 carbonatoms in the chain. Branched means that one or more lower alkyl groupssuch as methyl, ethyl or propyl, are attached to a linear alkyl chain.“Lower alkyl” means a group having about 1 to about 6 carbon atoms inthe chain which may be straight or branched. The term “substitutedalkyl” means that the alkyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of halo, alkyl, aryl,cycloalkyl, cyano, hydroxy, alkoxy, alkylthio, amino, —NH(alkyl),—NH(cycloalkyl), —N(alkyl)₂, —N(alkyl)₂, carboxy and —C(O)O-alkyl.Non-limiting examples of suitable alkyl groups include methyl, ethyl,n-propyl, isopropyl and t-butyl.

“Alkenyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon double bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkenyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 6 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkenyl chain. “Lower alkenyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. The term “substituted alkenyl” means that the alkenyl groupmay be substituted by one or more substituents which may be the same ordifferent, each substituent being independently selected from the groupconsisting of halo, alkyl, aryl, cycloalkyl, cyano, alkoxy and—S(alkyl). Non-limiting examples of suitable alkenyl groups includeethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, octenyland decenyl.

“Alkynyl” means an aliphatic hydrocarbon group containing at least onecarbon-carbon triple bond and which may be straight or branched andcomprising about 2 to about 15 carbon atoms in the chain. Preferredalkynyl groups have about 2 to about 12 carbon atoms in the chain; andmore preferably about 2 to about 4 carbon atoms in the chain. Branchedmeans that one or more lower alkyl groups such as methyl, ethyl orpropyl, are attached to a linear alkynyl chain. “Lower alkynyl” meansabout 2 to about 6 carbon atoms in the chain which may be straight orbranched. Non-limiting examples of suitable alkynyl groups includeethynyl, propynyl, 2-butynyl and 3-methylbutynyl. The term “substitutedalkynyl” means that the alkynyl group may be substituted by one or moresubstituents which may be the same or different, each substituent beingindependently selected from the group consisting of alkyl, aryl andcycloalkyl.

“Aryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 6 to about 14 carbon atoms, preferably about 6 to about10 carbon atoms. The aryl group can be optionally substituted with oneor more “ring system substituents” which may be the same or different,and are as defined herein. Non-limiting examples of suitable aryl groupsinclude phenyl and naphthyl.

“Heteroaryl” means an aromatic monocyclic or multicyclic ring systemcomprising about 5 to about 14 ring atoms, preferably about 5 to about10 ring atoms, in which one or more of the ring atoms is an elementother than carbon, for example nitrogen, oxygen or sulfur, alone or incombination. Preferred heteroaryls contain about 5 to about 6 ringatoms. The “heteroaryl” can be optionally substituted by one or more“ring system substituents” which may be the same or different, and areas defined herein. The prefix aza, oxa or thia before the heteroarylroot name means that at least a nitrogen, oxygen or sulfur atomrespectively, is present as a ring atom. A nitrogen atom of a heteroarylcan be optionally oxidized to the corresponding N-oxide. Non-limitingexamples of suitable heteroaryls include pyridyl, pyrazinyl, furanyl,thienyl, pyrimidinyl, pyridone (including N-substituted pyridones),isoxazolyl, isothiazolyl, oxazolyl, thiazolyl, pyrazolyl, furazanyl,pyrrolyl, pyrazolyl, triazolyl, 1,2,4-thiadiazolyl, pyrazinyl,pyridazinyl, quinoxalinyl, phthalazinyl, oxindolyl,imidazo[1,2-a]pyridinyl, imidazo[2,1-b]thiazolyl, benzofurazanyl,indolyl, azaindolyl, benzimidazolyl, benzothienyl, quinolinyl,imidazolyl, thienopyridyl, quinazolinyl, thienopyrimidyl,pyrrolopyridyl, imidazopyridyl, isoquinolinyl, benzoazaindolyl,1,2,4-triazinyl, benzothiazolyl and the like. The term “heteroaryl” alsorefers to partially saturated heteroaryl moieties such as, for example,tetrahydroisoquinolyl, tetrahydroquinolyl and the like.

“Aralkyl” or “arylalkyl” means an aryl-alkyl- group in which the aryland alkyl are as previously described. Preferred aralkyls comprise alower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude benzyl, 2-phenethyl and naphthalenylmethyl. The bond to theparent moiety is through the alkyl.

“Alkylaryl” means an alkyl-aryl- group in which the alkyl and aryl areas previously described. Preferred alkylaryls comprise a lower alkylgroup. Non-limiting example of a suitable alkylaryl group is tolyl. Thebond to the parent moiety is through the aryl.

“Cycloalkyl” means a non-aromatic mono- or multicyclic ring systemcomprising about 3 to about 10 carbon atoms, preferably about 5 to about10 carbon atoms. Preferred cycloalkyl rings contain about 5 to about 7ring atoms. The cycloalkyl can be optionally substituted with one ormore “ring system substituents” which may be the same or different, andare as defined above. Non-limiting examples of suitable monocycliccycloalkyls include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyland the like. Non-limiting examples of suitable multicyclic cycloalkylsinclude 1-decalinyl, norbornyl, adamantyl and the like, as well aspartially saturated species such as, for example, indanyl,tetrahydronaphthyl and the like.

“Halogen” or “halo” means fluorine, chlorine, bromine, or iodine.Preferred are fluorine, chlorine and bromine.

“Ring system substituent” means a substituent attached to an aromatic ornon-aromatic ring system which, for example, replaces an availablehydrogen on the ring system. Ring system substituents may be the same ordifferent, each being independently selected from the group consistingof alkyl, alkenyl, alkynyl, aryl, heteroaryl, aralkyl, alkylaryl,heteroaralkyl, heteroarylalkenyl, heteroarylalkynyl, alkylheteroaryl,hydroxy, hydroxyalkyl, alkoxy, aryloxy, aralkoxy, acyl, aroyl, halo,nitro, cyano, carboxy, alkoxycarbonyl, aryloxycarbonyl,aralkoxycarbonyl, alkylsulfonyl, arylsulfonyl, heteroarylsulfonyl,alkylthio, arylthio, heteroarylthio, aralkylthio, heteroaralkylthio,cycloalkyl, heterocyclyl, —C(═N—CN)—NH₂, —C(═NH)—NH₂, —C(═NH)—NH(alkyl),Y₁Y₂N—, Y₁Y₂N-alkyl-, Y₁Y₂NC(O)—, Y₁Y₂NSO₂— and —SO₂NY₁Y₂, wherein Y₁and Y₂ can be the same or different and are independently selected fromthe group consisting of hydrogen, alkyl, aryl, cycloalkyl, and aralkyl.“Ring system substituent” may also mean a single moiety whichsimultaneously replaces two available hydrogens on two adjacent carbonatoms (one H on each carbon) on a ring system. Examples of such moietyare methylene dioxy, ethylenedioxy, —C(CH₃)₂—and the like which formmoieties such as, for example:

“Heterocyclyl” means a non-aromatic saturated monocyclic or multicyclicring system comprising about 3 to about 10 ring atoms, preferably about5 to about 10 ring atoms, in which one or more of the atoms in the ringsystem is an element other than carbon, for example nitrogen, oxygen orsulfur, alone or in combination. There are no adjacent oxygen and/orsulfur atoms present in the ring system. Preferred heterocyclyls containabout 5 to about 6 ring atoms. The prefix aza, oxa or thia before theheterocyclyl root name means that at least a nitrogen, oxygen or sulfuratom respectively is present as a ring atom. Any —NH in a heterocyclylring may exist protected such as, for example, as an —N(Boc), —N(CBz),—N(Tos) group and the like; such protections are also considered part ofthis invention. The heterocyclyl can be optionally substituted by one ormore “ring system substituents” which may be the same or different, andare as defined herein. The nitrogen or sulfur atom of the heterocyclylcan be optionally oxidized to the corresponding N-oxide, S-oxide orS,S-dioxide. Non-limiting examples of suitable monocyclic heterocyclylrings include piperidyl, pyrrolidinyl, piperazinyl, morpholinyl,thiomorpholinyl, thiazolidinyl, 1,4-dioxanyl, tetrahydrofuranyl,tetrahydrothiophenyl, lactam, lactone, and the like.

It should be noted that in hetero-atom containing ring systems of thisinvention, there are no hydroxyl groups on carbon atoms adjacent to a N,O or S, as well as there are no N or S groups on carbon adjacent toanother heteroatom. Thus, for example, in the ring:

there is no —OH attached directly to carbons marked 2 and 5.

It should also be noted that tautomeric forms such as, for example, themoieties:

are considered equivalent in certain embodiments of this invention.

“Alkynylalkyl” means an alkynyl-alkyl- group in which the alkynyl andalkyl are as previously described. Preferred alkynylalkyls contain alower alkynyl and a lower alkyl group. The bond to the parent moiety isthrough the alkyl. Non-limiting examples of suitable alkynylalkyl groupsinclude propargylmethyl.

“Heteroaralkyl” means a heteroaryl-alkyl- group in which the heteroaryland alkyl are as previously described. Preferred heteroaralkyls containa lower alkyl group. Non-limiting examples of suitable aralkyl groupsinclude pyridylmethyl, and quinolin-3-ylmethyl. The bond to the parentmoiety is through the alkyl.

“Hydroxyalkyl” means a HO-alkyl- group in which alkyl is as previouslydefined. Preferred hydroxyalkyls contain lower alkyl. Non-limitingexamples of suitable hydroxyalkyl groups include hydroxymethyl and2-hydroxyethyl.

“Acyl” means an H—C(O)—, alkyl-C(O)— or cycloalkyl-C(O)—, group in whichthe various groups are as previously described. The bond to the parentmoiety is through the carbonyl. Preferred acyls contain a lower alkyl.Non-limiting examples of suitable acyl groups include formyl, acetyl andpropanoyl.

“Aroyl” means an aryl-C(O)— group in which the aryl group is aspreviously described. The bond to the parent moiety is through thecarbonyl. Non-limiting examples of suitable groups include benzoyl and1-naphthoyl.

“Alkoxy” means an alkyl-O— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkoxy groupsinclude methoxy, ethoxy, n-propoxy, isopropoxy and n-butoxy. The bond tothe parent moiety is through the ether oxygen.

“Aryloxy” means an aryl-O— group in which the aryl group is aspreviously described. Non-limiting examples of suitable aryloxy groupsinclude phenoxy and naphthoxy. The bond to the parent moiety is throughthe ether oxygen.

“Aralkyloxy” means an aralkyl-O— group in which the aralkyl group is aspreviously described. Non-limiting examples of suitable aralkyloxygroups include benzyloxy and 1- or 2-naphthalenemethoxy. The bond to theparent moiety is through the ether oxygen.

“Alkylthio” means an alkyl-S— group in which the alkyl group is aspreviously described. Non-limiting examples of suitable alkylthio groupsinclude methylthio and ethylthio. The bond to the parent moiety isthrough the sulfur.

“Arylthio” means an aryl-S— group in which the aryl group is aspreviously described. Non-limiting examples of suitable arylthio groupsinclude phenylthio and naphthylthio. The bond to the parent moiety isthrough the sulfur.

“Aralkylthio” means an aralkyl-S— group in which the aralkyl group is aspreviously described. Non-limiting example of a suitable aralkylthiogroup is benzylthio. The bond to the parent moiety is through thesulfur.

“Alkoxycarbonyl” means an alkyl-O—CO— group. Non-limiting examples ofsuitable alkoxycarbonyl groups include methoxycarbonyl andethoxycarbonyl. The bond to the parent moiety is through the carbonyl.

“Aryloxycarbonyl” means an aryl-O—C(O)— group. Non-limiting examples ofsuitable aryloxycarbonyl groups include phenoxycarbonyl andnaphthoxycarbonyl. The bond to the parent moiety is through thecarbonyl.

“Aralkoxycarbonyl” means an aralkyl-O—C(O)— group. Non-limiting exampleof a suitable aralkoxycarbonyl group is benzyloxycarbonyl. The bond tothe parent moiety is through the carbonyl.

“Alkylsulfonyl” means an alkyl-S(O₂)— group. Preferred groups are thosein which the alkyl group is lower alkyl. The bond to the parent moietyis through the sulfonyl.

“Arylsulfonyl” means an aryl-S(O₂)— group. The bond to the parent moietyis through the sulfonyl.

The term “substituted” means that one or more hydrogens on thedesignated atom is replaced with a selection from the indicated group,provided that the designated atom's normal valency under the existingcircumstances is not exceeded, and that the substitution results in astable compound. Combinations of substituents and/or variables arepermissible only if such combinations result in stable compounds. By“stable compound” or “stable structure” is meant a compound that issufficiently robust to survive isolation to a useful degree of purityfrom a reaction mixture, and formulation into an efficacious therapeuticagent.

The term “one or more” or “at least one”, when indicating the number ofsubstituents, compounds, combination agents and the like, refers to atleast one, and up to the maximum number of chemically and physicallypermissible, substituents, compounds, combination agents and the like,that are present or added, depending on the context. Such techniques andknowledge are well known within the skills of the concerned artisan.

The term “optionally substituted” means optional substitution with thespecified groups, radicals or moieties.

The term “isolated” or “in isolated form” for a compound refers to thephysical state of said compound after being isolated from a syntheticprocess or natural source or combination thereof. The term “purified” or“in purified form” for a compound refers to the physical state of saidcompound after being obtained from a purification process or processesdescribed herein or well known to the skilled artisan, in sufficientpurity to be characterizable by standard analytical techniques describedherein or well known to the skilled artisan.

It should also be noted that any carbon or heteroatom with unsatisfiedvalences in the text, schemes, examples and Tables herein is assumed tohave the hydrogen atom(s) to satisfy the valences.

When a functional group in a compound is termed “protected”, this meansthat the group is in modified form to preclude undesired side reactionsat the protected site when the compound is subjected to a reaction.Suitable protecting groups will be recognized by those with ordinaryskill in the art as well as by reference to standard textbooks such as,for example, T. W. Greene et al, Protective Groups in organic Synthesis(1991), Wiley, New York.

When any variable (e.g., aryl, heterocycle, R², etc.) occurs more thanone time in any constituent or in Formula 1, its definition on eachoccurrence is independent of its definition at every other occurrence.

As used herein, the term “composition” is intended to encompass aproduct comprising the specified ingredients in the specified amounts,as well as any product which results, directly or indirectly, fromcombination of the specified ingredients in the specified amounts.

Prodrugs and solvates of the compounds of the invention are alsocontemplated herein. The term “prodrug”, as employed herein, denotes acompound that is a drug precursor which, upon administration to asubject, undergoes chemical conversion by metabolic or chemicalprocesses to yield a compound of Formula 1 or a salt and/or solvatethereof. A discussion of prodrugs is provided in T. Higuchi and V.Stella, Pro-drugs as Novel Delivery Systems (1987) 14 of the A.C.S.Symposium Series, and in Bioreversible Carriers in Drug Design, (1987)Edward B. Roche, ed., American Pharmaceutical Association and PergamonPress, both of which are incorporated herein by reference thereto.

“Solvate” means a physical association of a compound of this inventionwith one or more solvent molecules. This physical association involvesvarying degrees of ionic and covalent bonding, including hydrogenbonding. In certain instances the solvate will be capable of isolation,for example when one or more solvent molecules are incorporated in thecrystal lattice of the crystalline solid. “Solvate” encompasses bothsolution-phase and isolatable solvates. Non-limiting examples ofsuitable solvates include ethanolates, methanolates, and the like.“Hydrate” is a solvate wherein the solvent molecule is H₂O.

“Effective amount” or “therapeutically effective amount” is meant todescribe an amount of compound or a composition of the present inventioneffective in inhibiting the CDK(s) and thus producing the desiredtherapeutic, ameliorative, inhibitory or preventative effect.

The compounds of Formula 1 can form salts which are also within thescope of this invention. Reference to a compound of Formula 1 herein isunderstood to include reference to salts thereof, unless otherwiseindicated. The term “salt(s)”, as employed herein, denotes acidic saltsformed with inorganic and/or organic acids, as well as basic saltsformed with inorganic and/or organic bases. In addition, when a compoundof Formula 1 contains both a basic moiety, such as, but not limited to apyridine or imidazole, and an acidic moiety, such as, but not limited toa carboxylic acid, zwitterions (“inner salts”) may be formed and areincluded within the term “salt(s)” as used herein. Pharmaceuticallyacceptable (i.e., non-toxic, physiologically acceptable) salts arepreferred, although other salts are also useful. Salts of the compoundsof the Formula 1 may be formed, for example, by reacting a compound ofFormula 1 with an amount of acid or base, such as an equivalent amount,in a medium such as one in which the salt precipitates or in an aqueousmedium followed by lyophilization.

Exemplary acid addition salts include acetates, ascorbates, benzoates,benzenesulfonates, bisulfates, borates, butyrates, citrates,camphorates, camphorsulfonates, fumarates, hydrochlorides,hydrobromides, hydroiodides, lactates, maleates, methanesulfonates,naphthalenesulfonates, nitrates, oxalates, phosphates, propionates,salicylates, succinates, sulfates, tartarates, thiocyanates,toluenesulfonates (also known as tosylates,) and the like. Additionally,acids which are generally considered suitable for the formation ofpharmaceutically useful salts from basic pharmaceutical compounds arediscussed, for example, by P. Stahl et al, Camille G. (eds.) Handbook ofPharmaceutical Salts. Properties, Selection and Use. (2002) Zurich:Wiley-VCH; S. Berge et al, Journal of Pharmaceutical Sciences (1977)66(1) 1–19; P. Gould, International J. of Pharmaceutics (1986) 33201–217; Anderson et al, The Practice of Medicinal Chemistry (1996),Academic Press, New York; and in The Orange Book (Food & DrugAdministration, Washington, D.C. on their website). These disclosuresare incorporated herein by reference thereto.

Exemplary basic salts include ammonium salts, alkali metal salts such assodium, lithium, and potassium salts, alkaline earth metal salts such ascalcium and magnesium salts, salts with organic bases (for example,organic amines) such as dicyclohexylamines, t-butyl amines, and saltswith amino acids such as arginine, lysine and the like. Basicnitrogen-containing groups may be quarternized with agents such as loweralkyl halides (e.g. methyl, ethyl, and butyl chlorides, bromides andiodides), dialkyl sulfates (e.g. dimethyl, diethyl, and dibutylsulfates), long chain halides (e.g. decyl, lauryl, and stearylchlorides, bromides and iodides), aralkyl halides (e.g. benzyl andphenethyl bromides), and others.

All such acid salts and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Pharmaceutically acceptable esters of the present compounds include thefollowing groups: (1) carboxylic acid esters obtained by esterificationof the hydroxy groups, in which the non-carbonyl moiety of thecarboxylic acid portion of the ester grouping is selected from straightor branched chain alkyl (for example, acetyl, n-propyl, t-butyl, orn-butyl), alkoxyalkyl (for example, methoxymethyl), aralkyl (forexample, benzyl), aryloxyalkyl (for example, phenoxymethyl), aryl (forexample, phenyl optionally substituted with, for example, halogen,C₁₋₄alkyl, or C₁₋₄alkoxy or amino); (2) sulfonate esters, such as alkyl-or aralkylsulfonyl (for example, methanesulfonyl); (3) amino acid esters(for example, L-valyl or L-isoleucyl); (4) phosphonate esters and (5)mono-, di- or triphosphate esters. The phosphate esters may be furtheresterified by, for example, a C₁₋₂₀ alcohol or reactive derivativethereof, or by a 2,3-di (C₆₋₂₄) acyl glycerol.

Compounds of Formula 1, and salts, solvates, esters and prodrugsthereof, may exist in their tautomeric form (for example, as an amide orimino ether). All such tautomeric forms are contemplated herein as partof the present invention.

All stereoisomers (for example, geometric isomers, optical isomers andthe like) of the present compounds (including those of the salts,solvates, esters and prodrugs of the compounds as well as the salts andsolvates of the prodrugs), such as those which may exist due toasymmetric carbons on various substituents, including enantiomeric forms(which may exist even in the absence of asymmetric carbons), rotamericforms, atropisomers, and diastereomeric forms, are contemplated withinthe scope of this invention, as are positional isomers (such as, forexample, 4-pyridyl and 3-pyridyl). Individual stereoisomers of thecompounds of the invention may, for example, be substantially free ofother isomers, or may be admixed, for example, as racemates or with allother, or other selected, stereoisomers. The chiral centers of thepresent invention can have the S or R configuration as defined by theIUPAC 1974 Recommendations. The use of the terms “salt”, “solvate”“prodrug” and the like, is intended to equally apply to the salt,solvate and prod rug of enantiomers, stereoisomers, rotamers, tautomers,positional isomers, racemates or prodrugs of the inventive compounds.

Polymorphic forms of the compounds of Formula I, and of the salts,solvates, esters and prodrugs of the compounds of Formula I, areintended to be included in the present invention.

It is to be understood that the utility of the compounds of Formula 1for the therapeutic applications discussed herein is applicable to eachcompound by itself or to the combination or combinations of one or morecompounds of Formula 1 as illustrated, for example, in the nextimmediate paragraph. The same understanding also applies topharmaceutical composition(s) comprising such compound or compounds andmethod(s) of treatment involving such compound or compounds.

The compounds according to the invention can have pharmacologicalproperties; in particular, the compounds of Formula 1 can be inhibitorsof HCV protease, each compound by itself or one or more compounds ofFormula 1 can be combined with one or more compounds selected fromwithin Formula 1. The compound(s) can be useful for treating diseasessuch as, for example, HCV, HIV, (AIDS, Acquired Immune DeficiencySyndrome), and related disorders, as well as for modulating the activityof hepatitis C virus (HCV) protease, preventing HCV, or ameliorating oneor more symptoms of hepatitis C.

The compounds of Formula 1 may be used for the manufacture of amedicament to treat disorders associated with the HCV protease, forexample, the method comprising bringing into intimate contact a compoundof Formula 1 and a pharmaceutically acceptable carrier.

In another embodiment, this invention provides pharmaceuticalcompositions comprising the inventive compound or compounds as an activeingredient. The pharmaceutical compositions generally additionallycomprise at least one pharmaceutically acceptable carrier diluent,excipient or carrier (collectively referred to herein as carriermaterials). Because of their HCV inhibitory activity, suchpharmaceutical compositions possess utility in treating hepatitis C andrelated disorders.

In yet another embodiment, the present invention discloses methods forpreparing pharmaceutical compositions comprising the inventive compoundsas an active ingredient. In the pharmaceutical compositions and methodsof the present invention, the active ingredients will typically beadministered in admixture with suitable carrier materials suitablyselected with respect to the intended form of administration, i.e. oraltablets, capsules (either solid-filled, semi-solid filled or liquidfilled), powders for constitution, oral gels, elixirs, dispersiblegranules, syrups, suspensions, and the like, and consistent withconventional pharmaceutical practices. For example, for oraladministration in the form of tablets or capsules, the active drugcomponent may be combined with any oral non-toxic pharmaceuticallyacceptable inert carrier, such as lactose, starch, sucrose, cellulose,magnesium stearate, dicalcium phosphate, calcium sulfate, talc,mannitol, ethyl alcohol (liquid forms) and the like. Moreover, whendesired or needed, suitable binders, lubricants, disintegrating agentsand coloring agents may also be incorporated in the mixture. Powders andtablets may be comprised of from about 5 to about 95 percent inventivecomposition.

Suitable binders include starch, gelatin, natural sugars, cornsweeteners, natural and synthetic gums such as acacia, sodium alginate,carboxymethylcellulose, polyethylene glycol and waxes. Among thelubricants there may be mentioned for use in these dosage forms, boricacid, sodium benzoate, sodium acetate, sodium chloride, and the like.Disintegrants include starch, methylcellulose, guar gum and the like.

Sweetening and flavoring agents and preservatives may also be includedwhere appropriate. Some of the terms noted above, namely disintegrants,diluents, lubricants, binders and the like, are discussed in more detailbelow.

Additionally, the compositions of the present invention may beformulated in sustained release form to provide the rate controlledrelease of any one or more of the components or active ingredients tooptimize the therapeutic effects, i.e. HCV inhibitory activity and thelike. Suitable dosage forms for sustained release include layeredtablets containing layers of varying disintegration rates or controlledrelease polymeric matrices impregnated with the active components andshaped in tablet form or capsules containing such impregnated orencapsulated porous polymeric matrices.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injections or addition of sweeteners and pacifiers fororal solutions, suspensions and emulsions. Liquid form preparations mayalso include solutions for intranasal administration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier such as inert compressed gas, e.g.nitrogen.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides such as cocoa butter is first melted, and theactive ingredient is dispersed homogeneously therein by stirring orsimilar mixing. The molten homogeneous mixture is then poured intoconvenient sized molds, allowed to cool and thereby solidify.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions may take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

The compounds of the invention may also be administered orally,intravenously, intranasally or subcutaneously.

The compounds of the invention may also comprise preparations which arein a unit dosage form. In such form, the preparation is subdivided intosuitably sized unit doses containing appropriate quantities of theactive components, e.g., an effective amount to achieve the desiredpurpose.

The quantity of the inventive active composition in a unit dose ofpreparation may be generally varied or adjusted from about 1.0 milligramto about 1,000 milligrams, preferably from about 1.0 to about 950milligrams, more preferably from about 1.0 to about 500 milligrams, andtypically from about 1 to about 250 milligrams, according to theparticular application. The actual dosage employed may be varieddepending upon the patient's age, sex, weight and severity of thecondition being treated. Such techniques are well known to those skilledin the art.

Generally, the human oral dosage form containing the active ingredientscan be administered 1 or 2 times per day. The amount and frequency ofthe administration will be regulated according to the judgment of theattending clinician. A generally recommended daily dosage regimen fororal administration may range from about 1.0 milligram to about 1,000milligrams per day, in single or divided doses.

Some useful terms are described below:

Capsule—refers to a special container or enclosure made of methylcellulose, polyvinyl alcohols, or denatured gelatins or starch forholding or containing compositions comprising the active ingredients.Hard shell capsules are typically made of blends of relatively high gelstrength bone and pork skin gelatins. The capsule itself may containsmall amounts of dyes, opaquing agents, plasticizers and preservatives.

Tablet—refers to a compressed or molded solid dosage form containing theactive ingredients with suitable diluents. The tablet can be prepared bycompression of mixtures or granulations obtained by wet granulation, drygranulation or by compaction.

Oral gel—refers to the active ingredients dispersed or solubilized in ahydrophillic semi-solid matrix.

Powder for constitution refers to powder blends containing the activeingredients and suitable diluents which can be suspended in water orjuices.

Diluent—refers to substances that usually make up the major portion ofthe composition or dosage form. Suitable diluents include sugars such aslactose, sucrose, mannitol and sorbitol; starches derived from wheat,corn, rice and potato; and celluloses such as microcrystallinecellulose. The amount of diluent in the composition can range from about10 to about 90% by weight of the total composition, preferably fromabout 25 to about 75%, more preferably from about 30 to about 60% byweight, even more preferably from about 12 to about 60%.

Disintegrant—refers to materials added to the composition to help itbreak apart (disintegrate) and release the medicaments. Suitabledisintegrants include starches; “cold water soluble” modified starchessuch as sodium carboxymethyl starch; natural and synthetic gums such aslocust bean, karaya, guar, tragacanth and agar; cellulose derivativessuch as methylcellulose and sodium carboxymethylcellulose;microcrystalline celluloses and cross-linked microcrystalline cellulosessuch as sodium croscarmellose; alginates such as alginic acid and sodiumalginate; clays such as bentonites; and effervescent mixtures. Theamount of disintegrant in the composition can range from about 2 toabout 15% by weight of the composition, more preferably from about 4 toabout 10% by weight.

Binder—refers to substances that bind or “glue” powders together andmake them cohesive by forming granules, thus serving as the “adhesive”in the formulation. Binders add cohesive strength already available inthe diluent or bulking agent. Suitable binders include sugars such assucrose; starches derived from wheat, corn rice and potato; natural gumssuch as acacia, gelatin and tragacanth; derivatives of seaweed such asalginic acid, sodium alginate and ammonium calcium alginate; cellulosicmaterials such as methylcellulose and sodium carboxymethylcellulose andhydroxypropylmethylcellulose; polyvinylpyrrolidone; and inorganics suchas magnesium aluminum silicate. The amount of binder in the compositioncan range from about 2 to about 20% by weight of the composition, morepreferably from about 3 to about 10% by weight, even more preferablyfrom about 3 to about 6% by weight.

Lubricant—refers to a substance added to the dosage form to enable thetablet, granules, etc. after it has been compressed, to release from themold or die by reducing friction or wear. Suitable lubricants includemetallic stearates such as magnesium stearate, calcium stearate orpotassium stearate; stearic acid; high melting point waxes; and watersoluble lubricants such as sodium chloride, sodium benzoate, sodiumacetate, sodium oleate, polyethylene glycols and d'l-leucine. Lubricantsare usually added at the very last step before compression, since theymust be present on the surfaces of the granules and in between them andthe parts of the tablet press. The amount of lubricant in thecomposition can range from about 0.2 to about 5% by weight of thecomposition, preferably from about 0.5 to about 2%, more preferably fromabout 0.3 to about 1.5% by weight.

Glident—material that prevents caking and improve the flowcharacteristics of granulations, so that flow is smooth and uniform.Suitable glidents include silicon dioxide and talc. The amount ofglident in the composition can range from about 0.1% to about 5% byweight of the total composition, preferably from about 0.5 to about 2%by weight.

Coloring agents—excipients that provide coloration to the composition orthe dosage form. Such excipients can include food grade dyes and foodgrade dyes adsorbed onto a suitable adsorbent such as clay or aluminumoxide. The amount of the coloring agent can vary from about 0.1 to about5% by weight of the composition, preferably from about 0.1 to about 1%.

Bioavailability—refers to the rate and extent to which the active drugingredient or therapeutic moiety is absorbed into the systemiccirculation from an administered dosage form as compared to a standardor control.

Conventional methods for preparing tablets are known. Such methodsinclude dry methods such as direct compression and compression ofgranulation produced by compaction, or wet methods or other specialprocedures. Conventional methods for making other forms foradministration such as, for example, capsules, suppositories and thelike are also well known.

Another embodiment of the invention discloses the use of the inventivecompounds or pharmaceutical compositions disclosed above for treatmentof diseases such as, for example, hepatitis C and the like. The methodcomprises administering a therapeutically effective amount of theinventive compound or pharmaceutical composition to a patient havingsuch a disease or diseases and in need of such a treatment.

In yet another embodiment, the compounds of the invention may be usedfor the treatment of HCV in humans in monotherapy mode or in acombination therapy (e.g., dual combination, triple combination etc.)mode such as, for example, in combination with antiviral and/orimmunomodulatory agents. Examples of such antiviral and/orimmunomodulatory agents include Ribavirin (from Schering-PloughCorporation, Madison, N.J.) and Levovirin™ (from ICN Pharmaceuticals,Costa Mesa, Calif.), VP50406™ (from Viropharma, Incorporated, Exton,Pa.), ISIS14803™ (from ISIS Pharmaceuticals, Carlsbad, Calif.),Heptazyme™ (from Ribozyme Pharmaceuticals, Boulder, Colo.), VX497™ (fromVertex Pharmaceuticals, Cambridge, Mass.), Thymosin™ (from SciClonePharmaceuticals, San Mateo, Calif.), Maxamine™ (Maxim Pharmaceuticals,San Diego, Calif.), mycophenolate mofetil (from Hoffman-LaRoche, Nutley,N.J.), interferon (such as, for example, interferon-alpha,PEG-interferon alpha conjugates) and the like. “PEG-interferon alphaconjugates” are interferon alpha molecules covalently attached to a PEGmolecule. Illustrative PEG-interferon alpha conjugates includeinterferon alpha-2a (Roferon™, from Hoffman La-Roche, Nutley, N.J.) inthe form of pegylated interferon alpha-2a (e.g., as sold under the tradename Pegasys™), interferon alpha-2b (Intron™, from Schering-PloughCorporation) in the form of pegylated interferon alpha-2b (e.g., as soldunder the trade name PEG-Intron™), interferon alpha-2c (Berofor Alpha™,from Boehringer Ingelheim, Ingelheim, Germany) or consensus interferonas defined by determination of a consensus sequence of naturallyoccurring interferon alphas (Infergen™, from Amgen, Thousand Oaks,Calif.).

As stated earlier, the invention includes tautomers, rotamers,enantiomers and other stereoisomers of the inventive compounds also.Thus, as one skilled in the art appreciates, some of the inventivecompounds may exist in suitable isomeric forms. Such variations arecontemplated to be within the scope of the invention.

Another embodiment of the invention discloses a method of making thecompounds disclosed herein. The compounds may be prepared by severaltechniques known in the art. Illustrative procedures are outlined in thefollowing reaction schemes. The illustrations should not be construed tolimit the scope of the invention which is defined in the appendedclaims. Alternative mechanistic pathways and analogous structures willbe apparent to those skilled in the art.

It is to be understood that while the following illustrative schemesdescribe the preparation of a few representative inventive compounds,suitable substitution of any of both the natural and unnatural aminoacids will result in the formation of the desired compounds based onsuch substitution. Such variations are contemplated to be within thescope of the invention.

For the procedures described below, the following abbreviations areused:

ABBREVIATIONS

Abbreviations which are used in the descriptions of the schemes,preparations and the examples that follow are:

-   THF: Tetrahydrofuran-   DMF: N,N-Dimethylformamide-   EtOAc: Ethyl acetate-   AcOH: Acetic acid-   HOOBt: 3-Hydroxy-1,2,3-benzotriazin-4(3H)-one-   EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride-   NMM: N-Methylmorpholine-   ADDP: 1,1′-(Azodicarbobyl)dipiperidine-   DEAD: Diethylazodicarboxylate-   DIAD: Diisopropylazodicarboxylate-   MeOH: Methanol-   EtOH: Ethanol-   Et₂O: Diethyl ether-   DMSO: Dimethylsulfoxide-   HOBt: N-Hydroxybenzotriazole-   PyBrOP: Bromo-tris-pyrrolidinophosphonium hexafluorophosphate-   DCM: Dichloromethane-   DCC: 1,3-Dicyclohexylcarbodiimide-   TEMPO: 2,2,6,6-Tetramethyl-1-piperidinyloxy-   Phg: Phenylglycine-   Chg: Cyclohexylglycine-   Bn: Benzyl-   Bz: Benzyl-   Et: Ethyl-   Ph: Phenyl-   iBoc: isobutoxycarbonyl-   iPr: isopropyl-   ^(t)Bu or Bu^(t): tert-Butyl-   Boc: tert-Butyloxycarbonyl-   Cbz: Benzyloxycarbonyl-   Cp: Cylcopentyidienyl-   Ts: p-toluenesulfonyl-   Me: Methyl-   Ms or Mesyl: Methane sulfonyl-   HATU: O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium-   hexafluorophosphate-   DMAP: 4-N,N-Dimethylaminopyridine-   Bop: Benzotriazol-1-yl-oxy-tris(dimethylamino)hexafluorophosphate-   PCC: Pyridiniumchlorochromate-   DIBAL-H: diisopropyl aluminum hydride-   rt or RT: Room temperature-   quant.: Quantitative yield-   h or hr: hour-   min: minute-   TFA: Trifluoroacetic acid

General Schemes for Preparation of Target Compounds

Compounds of the present invention were synthesized using the generalschemes (Methods A–E) described below.

Method A:

Deprotection of the N-Boc functionality of 1.01 under acidic conditionsprovided the hydrochloride salt 1.02 which was subsequently coupled withN-Boc-tert-leucine under peptide coupling methodology to afford 1.03.N-Boc deprotection followed by treatment with appropriate isocyanategave the urea 1.05. Hydrolysis of the methyl ester provided the acid1.06. Peptide coupling of the acid 1.06 with the appropriateP₁—P′primary amide moiety afforded the hydroxylamide 1.07. Oxidation(Moffatt or related process—T. T. Tidwell, Synthesis, 1990, 857; orDess-Martin's periodinane (J. Org. Chem., 1983, 48, 4155) resulted inthe target compound 1.08.

Method B

Peptide coupling of the acid 1.06 with the appropriate P₁—P′secondaryamide moiety afforded the hydroxylamide 1.09. Oxidation (Moffatt orDess-Martin's) resulted in the target compound 1.10.

Method C

In another variation, peptide coupling of the N-Boc-P₂—P₃-acid 1.17 withthe appropriate P₁—P′amide moiety afforded the hydroxylamide 1.11.Oxidation (Moffatt or Dess-Martin's) resulted in the keto amide 1.12.Deprotection of the N-Boc functionality gave the hydrochloride salt1.13. Treatment with a suitable isocyanate (or isocyanate equivalent)resulted in the target compound 1.14.

Method D

In yet another variation, the hydrochloride salt 1.13 was converted tothe 4-nitrophenyl carbamate 1.15 by reaction with 4-nitrophenylchloroformate. Subsequent treatment with an amine (or aminehydrochloride salt) of choice provided the target compound 1.14.

Method E

In yet another variation, the dipeptide hydrochloride salt 1.03 wasconverted to the 4-nitrophenyl carbamate as described above. Treatmentwith an amine (or amine hydrochloride salt) of choice provided the ureaderivative 1.05. Hydrolysis and further elaboration as described inMethods A/B provided the target compounds 1.14.

Preparation of Intermediates

Preparation of Intermediates 10.11 and 10.12:

A stirred solution of ketimine 10.01 (50 g, 187.1 mmol) under N₂ in dryTHF (400 mL) was cooled to −78°C. and treated with 1 M solution ofK-^(t)BuO (220 mL, 1.15 equiv.) in THF. The reaction mixture was warmedto 0°C. and stirred for 1 h and treated with bromomethyl cyclobutane (28mL, 249 mmol). The reaction mixture was stirred at room temperature for48 h and concentrated in vacuo. The residue was dissolved in Et₂O (300mL) and treated with aq. HCl (2 M, 300 mL) The resulting solution wasstirred at room temperature for 5 h and extracted with Et₂O (1 L). Theaqueous layer was made basic to pH˜12–14 with NaOH (50% aq.) andextracted with CH₂Cl₂ (3×300 mL). The combined organic layers were dried(MgSO₄), filtered, and concentrated to give the pure amine (10.02, 18 g)as a colorless oil.

A solution of the amine 10.02 (18 g, 105.2 mmol) at 0°C. in CH₂Cl₂ (350mL) was treated with di-tert-butyldicarbonate (23 g, 105.4 mmol) andstirred at rt. for 12 h. After the completion of the reaction (TLC), thereaction mixture was concentrated in vacuo and the residue was dissolvedin THF/H₂O (200 ml, 1:1) and treated with LiOH.H₂O (6.5 g, 158.5 mmol)and stirred at room temperature for 3 h. The reaction mixture wasconcentrated and the basic aqueous layer was extracted with Et₂O. Theaqueous layer was acidified with conc. HCl to pH˜1–2 and extracted withCH₂Cl₂. The combined organic layers were dried (MgSO₄), filtered, andconcentrated in vacuo to yield 10.03 as a colorless viscous oil whichwas used for the next step without any further purification.

A solution of the acid 10.03 (15.0 g, 62 mmol) in CH₂Cl₂ (250 mL) wastreated with BOP reagent (41.1 g, 93 mmol), N-methyl morpholine (27 mL),N,O-dimethyl hydroxylamine hydrochloride (9.07 g, 93 mmol) and stirredovernight at rt. The reaction mixture was diluted with 1 N aq. HCl (250mL), and the layers were separated and the aqueous layer was extractedwith CH₂Cl₂ (3×300 ml). The combined organic layers were dried (MgSO₄),filtered and concentrated in vacuo and purified by chromatography (SiO₂,EtOAc/Hex 2:3) to yield the amide 10.04 (15.0 g) as a colorless solid.

A solution of the amide 10.04 (15 g, 52.1 mmol) in dry THF (200 mL) wastreated dropwise with a solution of LiAlH₄ (1M, 93 mL, 93 mmol) at 0° C.The reaction mixture was stirred at room temperature for 1 h andcarefully quenched at 0° C. with a solution of KHSO₄ (10% aq.) andstirred for 0.5 h. The reaction mixture was diluted with aq. HCl (1 M,150 mL) and extracted with CH₂Cl₂ (3×200 mL), The combined organiclayers were washed with aq. HCl (1 M), saturated NaHCO₃, brine, anddried (MgSO₄). The mixture was filtered and concentrated in vacuo toyield 10.05 as a viscous colorless oil (14 g).

A solution of the aldehyde 10.05 (14 g, 61.6 mmol) in CH₂Cl₂ (50 mL),was treated with Et₃N (10.73 mL, 74.4 mmol), and acetone cyanohydrin(10.86 g, 127.57 mmol) and stirred at room temperature for 24 hrs. Thereaction mixture was concentrated in vacuo and diluted with aq. HCl (1M, 200 mL) and extracted into CH₂Cl₂ (3×200 mL). The combined organiclayer were washed with H₂O, brine, dried (MgSO₄), filtered, concentratedin vacuo and purified by chromatography (SiO₂, EtOAc/Hex 1:4) to yield10.06 (10.3 g) as a colorless liquid

Methanol saturated with HCl*, prepared by bubbling HCl gas through CH₃OH(700 ml) at 0° C., was treated with the cyanohydrin 10.06 and heated toreflux for 24 h. The reaction was concentrated in vacuo to yield 10.07,which was used in the next step without purification.

* Alternatively 6M HCl prepared by addition of AcCl to dry methanol canalso be used.

A solution of the amine hydrochloride 10.07 in CH₂Cl₂ (200 mL) wastreated with Et₃N (45.0 mL, 315 mmol) and Boc₂O (45.7 g, 209 mmol) at−78° C. The reaction mixture was then stirred at room temperatureovernight and diluted with HCl (2 M, 200 mL) and extracted into CH₂Cl₂.The combined organic layer were dried (MgSO₄) filtered, concentrated invacuo and purified by chromatography (EtOAc/Hex 1:4) to yield hydroxyester 10.08.

A solution of methyl ester 10.08 (3 g, 10.5 mmol) in THF/H₂O (1:1) wastreated with LiOH.H₂O (645 mg, 15.75 mmol) and stirred at rt. for 2 h.The reaction mixture was acidified with aq HCl (1 M, 15 mL) andconcentrated in vacuo. The residue was dried in vacuum to afford 10.09in quantitative yield.

A solution of the acid 10.09 (from above) in CH₂Cl₂ (50 mL) and DMF (25mL) was treated with NH₄Cl (2.94 g, 55.5 mmol), EDCI (3.15 g, 16.5mmol), HOOBt (2.69 g, 16.5 mmol), and NMM (4.4 g, 44 mmol). The reactionmixture was stirred at room temperature for 3 d. The solvents wereremoved under vacuo and the residue was diluted with aq. HCl (250 mL)and extracted with CH₂Cl₂. The combined organic layers were washed withaq. Sat'd. NaHCO₃, dried (MgSO₄) filtered concentrated in vacuo toobtain 10.10, which was used as it was in the following steps.(Alternatively 10.10 can also be obtained directly by the reaction of10.06 (4.5 g, 17.7 mmol) with aq. H₂O₂ (10 mL), LiOH.H₂O (820 mg, 20.8mmol) at 0° C. in 50 mL of CH₃OH for 0.5 h.)

A solution of 10.10 obtained in the previous step was dissolved in 4 NHCl in dioxane and stirred at rt. for 2 h. The reaction mixture wasconcentrated in vacuo to give the intermediate 10.11 as a solid, whichwas used without further purification.

The required intermediate 10.12 was obtained from compound 10.09 usingessentially the procedures described above in Steps 9, 10 withappropriate reagents.

Preparation of Intermediate 11.01

To a solution of 4-pentyn-1-ol, 11.02 (4.15 g; Aldrich) was addedDess-Martin Periodinane (30.25 g; Aldrich) and the resulting mixture wasstirred for 45 min. before the addition of(tert-Butoxycarbonylmethylene)triphenylphosphorane (26.75 g; Aldrich).The resulting dark reaction was stirred overnight, diluted with EtOAc),washed with aq. sodium sulfite. sat. aq. NaHCO3, water, brine and dried.The volatiles were removed under reduced pressure and the residue waspurified by silica gel column chromatography using 1% EtOAc in hexanesas eluent to give the desired compound, 11.03 (3.92 g). Some impurefractions were also obtained but set aside at this time.

Using the alkene 11.03 (1.9 g) in n-propanol (20 ml; Aldrich)), benzylcarbamate (4.95 g; Aldrich) in n-propanol (40 ml), NaOH (1.29 g) inwater (79 ml), tert-butyl hypochlorite (3.7 ml), (DHQ)₂PHAL (0.423 g;Aldrich)) in n-propanol (37.5 ml), and potassium osmate:dehydrate(0.1544 g; Aldrich) and the procedure set forth in Angew. Chem. Int. Ed.Engl (1998), 35, (23/24), pp. 2813–7, gave a crude product which waspurified by silica gel column chromatography using EtOAc:Hexanes (1:5)to give the desired amino alcohol 11.04 (1.37 g, 37%) as a white solid.

To the ester 11.04 (0.700 g) was added 4M HCl in dioxane (20 ml;Aldrich) and the resulting mixture was allowed to stand at roomtemperature overnight. The volatiles were removed under reduced pressureto give the acid 11.05 (0.621 g) as a white solid.

BOP reagent (3.65 g; Sigma) followed by triethylamine (3.45 ml) wereadded to a dichloromethane (20 ml) solution of the carboxylic acid 11.05(2.00 g) and allyl amine (0.616 ml) at room temperature and theresulting mixture was stirred overnight. The reaction mixture waspartitioned between EtOAc and 10% aq. HCl. The organic phase wasseparated, washed with sat. aq. sodium bicarbonate, water, dried(magnesium sulfate). The crude reaction product was purified by silicagel column chromatography using (EtOAc:Hexanes; 70:30) as eluent toprovide the desired amide 11.01 (1.73 g) as a viscous yellow oil.

Preparation of Intermediates 12.03 and 12.04

Compound 12.01 was converted to the required material 12.02 usingessentially the procedures described for Intermediate 10.11, Steps 3–8.

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.11, Steps 9,10.

Compound 12.02 was converted to the required intermediate 12.03 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 13.01

To a stirred solution of 1-nitrobutane, 13.02 (16.5 g, 0.16 mol) andglyoxylic acid in H₂O (28.1 g, 0.305 mol) and MeOH (122 mL) at 0° C.–5°C., was added dropwise triethylamine (93 mL, 0.667 mol) over 2 hrs. Thesolution was warmed to room temperature, stirred overnight andconcentrated to dryness to give an oil. The oil was then dissolved inH₂O and acidified to pH=1 with 10% HCl, followed by extraction withEtOAc. The combined organic solution was washed with brine, dried overNa₂SO₄, filtered and concentrated to dryness to give the product 13.03(28.1 g, 99% yield).

To a stirred solution of compound 13.03 (240 g, 1.35 mol) in acetic acid(1.25 L) was added 10% Pd/C (37 g). The resulting solution washydrogenated at 59 psi for 3 hrs and then at 60 psi overnight. Theacetic acid was then evaporated and azeotroped 3 times with toluene,then triturated with MeOH and ether. The solution was then filtered andazeotroped twice with toluene to afford 13.04 as an off white solid (131g, 0.891 mol, 66%).

To a stirred solution of the amino acid 13.04 (2.0 g, 13.6 mmol) indioxane (10 mL) and H₂O (5 mL) at 0° C., was added 1N NaOH solution (4.3mL, 14.0 mmol). The resulting solution was stirred for 10 minutes,followed by addition of di-t-butyldicarbonate (0.110 g, 14.0 mmol) andstirred at 0° C. for 15 minutes. The solution was then warmed to roomtemperature, stirred for 45 minutes and kept at refrigerator overnightand concentrated to dryness to give a crude material. To the solution ofthis crude material in EtOAc (100 mL) and ice, was added KHSO₄ (3.36 g)and H₂O (32 mL) and stirred for 4–6 minutes. The organic layer was thenseparated and the aqueous layer was extracted twice with EtOAc and thecombined organic layer was washed with water, brine, dried over Na₂SO₄,filtered and concentrated to dryness to give the product 13.05 as aclear gum (3.0 g, 89% yield).

Compound 13.05 was converted to the required intermediate 13.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 14.01

Compound 14.02 was converted to the required material 14.03 usingessentially the procedures described for Intermediate 13.01, Steps 1–3.

Compound 14.03 was converted to the required intermediate 14.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 15.01

To a suspension of silver nitrite (9 g, 58.5 mmol) in diethyl ether (25mL) at 0° C. was added a solution of 4-iodo-1,1,1-trifluorobutane, 15.02(10 g, 42.0 mmol) in diethyl ether (25 mL) slowly through an additionfunnel (approx. 15 min). The resulting mixture was vigorously stirred at0° C. and warmed to rt. After 50 h, the solid material was filtered offthrough a celite pad. The resulting diethyl ether solution wasconcentrated in vacuo to give 15.03 as colorless oil, which was usedwithout further purification.

Compound 15.03 was converted to the required material 15.04 usingessentially the procedures described for Intermediate 13.01, Steps 1–3.

Compound 15.04 was converted to the required intermediate 15.01 usingessentially the procedures described for Intermediate 10.12, Step 11.

Preparation of Intermediate 16.01

The acid 16.02 (Winkler, D.; Burger, K., Synthesis, 1996, 1419) isprocessed as described above (preparation of Intermediate 10.12) to givethe expected intermediate 16.01.

Preparation of Intermediate 20.01

The amino ester 20.01 was prepared following the method of R. Zhang andJ. S. Madalengoitia (J. Org. Chem. 1999, 64, 330), with the exceptionthat the Boc group was cleaved by the reaction of the Boc-protectedamino acid with methanolic HCl (4M HCl in dioxane was also employed forthe deprotection). In a variation of the reported synthesis, thesulfonium ylide was replaced with the corresponding phosphonium ylide.

Preparation of Intermediate 20.04

A solution of commercial amino acid Boc-Chg-OH, 20.02 (Senn chemicals,6.64 g, 24.1 mmol) and amine hydrochloride 20.01 (4.5 g, 22 mmol) inCH₂Cl₂ (100 mL) at 0° C. was treated with BOP reagent and stirred at rt.for 15 h. The reaction mixture was concentrated in vacuo, then it wasdiluted with aq. 1 M HCl and extracted into EtOAc (3×200 mL). Thecombined organic layers were washed with sat'd. NaHCO₃ (200 mL), dried(MgSO₄), filtered and concentrated in vacuo, and chromatographed (SiO₂,EtOAc/Hex 3:7) to obtain 20.03 (6.0 g) as a colorless solid.

A solution of methyl ester 20.03 (4.0 g, 9.79 mmol) in THF/H₂O (1:1) wastreated with LiOH.H₂O (401 mg, 9.79 mmol) and stirred at rt. for 3 h.The reaction mixture was acidified with aq. HCl and concentrated invacuo to obtain the required intermediate, free acid 20.04.

Preparation of Intermediate 20.08

A solution of Boc-tert-Leu 20.05 (Fluka, 5.0 g 21.6 mmol) in dryCH₂Cl₂/DMF (50 mL, 1:1) was cooled to 0° C. and treated with the aminesalt 20.01 (5.3 g, 25.7 mmol), NMM (6.5 g, 64.8 mmol) and BOP reagent(11.6 g, 25.7 mmol). The reaction was stirred at rt. for 24 h, dilutedwith aq. HCl (1 M) and extracted with CH₂Cl₂. The combined organiclayers were washed with HCl (aq, 1 M), sat'd. NaHCO₃, brine, dried(MgSO₄), filtered and concentrated in vacuo and purified bychromatography (SiO2, Acetone/Hexane 1:5) to yield 20.06 as a colorlesssolid.

A solution of methyl ester 20.06 (4.0 g, 10.46 mmol) was dissolved in 4MHCl in dioxane and stirred at rt. for 3 h. The reaction mixture wasconcentrated in vacuo to obtain the amine hydrochloride salt, 20.07which was used without purification.

A solution of the amine salt 20.07 (840 mg, 2.64 mmol) in THF (14mL)/acetonitrile (2 mL) was cooled to 0° C. 4-Nitrophenylchloroformate(800 mg, 3.96 mmol) was added followed by pyridine (0.64 mL, 7.92 mmol).The reaction was slowly warmed to room temperature over 3 hrs when TLCindicated reaction completion. Diethyl ether (50 mL) was added and theresulting precipitate was filtered off. The filtrate was washed withsaturated ammonium chloride solution (1×), brine (1×), dried (Na₂SO₄)and concentrated. The residue was purified by flash chromatography using20/80 EtOAc/hexanes which afforded 1.15 g of the required intermediate20.08.

Preparation of Intermediate 21.01

To a stirred solution of N-Boc-3,4-dehydroproline 21.02 (5.0 g, 23.5mmol), di-tert-butyl dicarbonate (7.5 g, 34.4 mmol), and4-N,N-dimethylaminopyridine (0.40 g, 3.33 mmol) in acetonitrile (100 mL)at room temperature was added triethylamine (5.0 mL, 35.6 mmol). Theresulting solution was stirred at this temperature for 18 h before itwas concentrated in vacuo. The dark brown residue was purified by flashcolumn chromatography eluting with 10–25% EtOAc/hexane to give theproduct 21.03 as a pale yellow oil (5.29 g, 84%).

To a stirred solution of the dehydroproline derivative 21.03 (10.1 g,37.4 mmol), benzyltriethylammonium chloride (1.60 g, 7.02 mmol) inchloroform (120 mL) at room temperature was added 50% aqueous sodiumhydroxide (120 g). After vigorously stirred at this temperature for 24h, the dark mixture was diluted with CH₂Cl₂ (200 mL) and diethyl ether(600 mL). After the layers were separated, the aqueous solution wasextracted with CH₂Cl₂/Et₂O (1:2, 3×600 mL). The organic solution wasdried (MgSO₄) and concentrated. The residue was purified by flash columnchromatography using 5–20% EtOAc/hexane to afford 9.34 g (71%) of 21.04as an off-white solid.

The solution of 21.04 (9.34 g, 26.5 mmol) in CH₂Cl₂ (25 mL) and CF₃CO₂H(50 mL) was stirred at room temperature for 4.5 h before it wasconcentrated in vacuo to give a brown residue, 21.05 which was used inStep 4 without further purification.

Concentrated hydrochloric acid (4.5 mL) was added to a solution of theresidue 21.05 from Step 3 in methanol (70 mL) and the resulting mixturewas warmed to 65° C. in an oil bath. After 18 h, the mixture wasconcentrated in vacuo to give a brown oil 21.01, which was used furtherwithout purification.

Preparation of Intermediate 22.01

Potassium bis(trimethylsilyl)amide (158 ml of a 0.5M solution intoluene; 79 mmol) was added to a stirred suspension ofcyclopropyltriphenyl-phosphonium bromide (33.12 g; 86.4 mmol) inanhydrous tetrahydrofuran (130 ml) and the resulting orange mixture wasstirred under an atmosphere of nitrogen at room temperature for a periodof 1 h., before the addition of the aldehyde 22.02 (9.689; 42.2 mmol) inTHF (8 ml). The reaction was then refluxed under an atmosphere ofnitrogen for a period of 2 h. After cooling, methanol, diethyl ether andRochelles salt were added. The organic phase was separated, washed withbrine, dried and concentrated under reduced pressure. The crude reactionproduct was purified by silica gel column chromatography usingEtOAc-hexane (1:99) to EtOAc-hexane (5:95) to provide the alkene 22.03(8.47 g) as a yellow oil.

A solution of 1M HCl in MeOH/MeOAc was prepared by adding 14.2 ml ofacetyl chloride dropwise into cold methanol and diluting the resultingsolution to 200 ml at room temperature. The carbamate 22.03 (9.49 g;37.5 mmol) was dissolved in methanol (12 ml) and added to 1M HCl inMeOH/MeOAc (150 ml) while cooled in an ice bath. The resulting mixturewas maintained at this temperature for 1 h., then the ice bath wasremoved and stirring continued overnight at room temperature. Thevolatiles were removed under reduced pressure to yield a yellow oilwhich was used in the next step without purification.

The yellow oil was dissolved in a mixture of THF (30 ml) and MeOH (20ml) and treated with triethylamine (15 ml; 108 mmol) until the solutionwas pH=9–10. After placing in an ice bath, the mixture was treated withN-Boc-Gly-OSu (11.229; 41 mmol). The ice bath was withdrawn and thereaction stirred at room temp. for 1 h. The volatiles were removed underreduced pressure and the residue was purified by silica gel columnchromatography using methanol (1–3%) in dichloromethane providing thedesired amide 22.04 (9.09 g).

The alcohol 22.04 (9.09 g; 33.6 mmol) was dissolved in acetone (118.5ml) and treated with 2,2-dimethoxypropane (37.4 ml; 304 mmol) andBF₃:Et₂O (0.32 ml; 2.6 mmol) and the resulting mixture was stirred atroom temperature for a period of 5.5 h The reaction solution was treatedwith a few drops of triethylamine and the volatiles were removed underreduced pressure. The residue was purified by silica gel columnchromatography using 5–25% EtOAc in hexanes to provide the N,O-acetal22.05 (8.85 g).

The carbamate 22.05 (8.81 g; 28.4 mmol) was dissolved in acetonitrile(45 ml) and the solution was cooled to −40° C. under an atmosphere ofnitrogen. Pyridine (6.9 ml; 85.3 mmol) followed by nitrosiumtetrafluoroborate (6.63 g; 56.8 mmol) were added and the resultingreaction mixture maintained below 0° C. until TLC indicated that nostarting material remained (approx. 2.25 h.). Pyrrolidine (20 ml; 240mmol) was added and the cooling bath was withdrawn and stirring wascontinued at room temperature for 1 h. and then the volatiles wereremoved under reduced pressure. The residue was quickly passed through apad of silica gel to provide a yellow oil.

The yellow oil was dissolved in anhydrous benzene (220 ml) and palladiumacetate (0.317 g; 1.41 mmol) was added before heating the resultingmixture to reflux, under an atmosphere of nitrogen for a period of 1.5h. After cooling, the volatiles were removed under reduced pressure andthe dark residue was purified by silica gel column chromatography usingEtOAc-hexane (1:4) to provide the I) the trans-pyrrolidinone 22.06 (1.94g) followed by ii) the cis-pyrrolidinone 22.07 (1.97 g).

Freshly prepared 1M HCl in MeOAc/MeOH (10 ml; as described above) wasadded to the N,O-acetal 22.06 and stirred at room temperature for 1 h.The solvent was removed under reduced pressure and the residue waspurified by silica gel column chromatography using 0–4% MeOH indichloromethane as eluent to provide the desired alcohol 22.08 (1.42 g),a yellow oil.

To a solution of the lactam 22.08 (1.29 g; 8.44 mmol) in anhydroustetrahydrofuran (55 ml) was added lithium aluminum hydride (2.40 g; 63.2mmol) and the resulting mixture was refluxed for 8 h. After cooling,water, followed by 15% aq. NaOH were added and the resulting mixture wasfiltered through celite and the solid was washed thoroughly with THF andMeOH. The solvent was removed under reduced pressure and the residueredissolved in dichloromethane, dried and concentrated under reducedpressure to provide the pyrrolidine, used without purification.

Hunigs base (4.5 ml; 25.8 mmol) was added to a mixture ofN-Boc-L-tert-Leu-OH (1.76 g; 7.6 mmol), The crude pyrrolidine and HATU(2.89 g; 7.6 mmol) in anhydrous dichloromethane (50 ml) at −60° C.,under an atmosphere of nitrogen. The resulting reaction was allowed tocome to room temperature slowly, overnight. EtOAc was added and theyellow solution was washed with dil. aq. HCl, sat. aq. sodiumbicarbonate, water, brine. The organic layer was dried and concentratedunder reduced pressure. The residue was purified by silica gel columnchromatography using EtOAc:hexanes (1:3) to give the desired amide 22.09(2.00 g).

The alcohol 22.09 (2.00 g; 5.67 mmol) was dissolved in acetone (116 ml)and cooled in an ice bath for 10 min. This solution was then added to acooled Jones reagent (14.2 ml; approx 2 mmol/ml) and the resultingmixture was stirred at 5 C. for 0.5 h and the cooling bath was removed.The reaction was stirred for a further 2 h. at room temp., before addingto sodium sulfate (28.54 g), celite (15 g) in EtOAc (100 ml).Isopropanol (15 ml) was added after 1 min and then stirred for a further10 min. and filtered. The filtrate was concentrated under reducedpressure, providing a brown oil which was dissolved in EtOAc. Thissolution was washed with water, 3% aq. citric acid, brine, dried andconcentrated to provide the desired carboxylic acid 22.01 (1.64 g) as awhite solid.

Preparation of Intermediate 23.01

To the mixture of ester 23.02 (6.0 g) and molecular sieve (5.2 g) inanhydrous methylene chloride (35 mL) was added pyrrolidine (5.7 mL,66.36 mmoL). The resulting brown slurry was stirred at room temperatureunder N₂ for 24 h, filtered and washed with anhydrous CH₃CN. Thecombined filtrate was concentrated to yield the desired product, 23.03.

To a solution of the product 23.03 from proceeding step in CH₃CN (35 mL)was added anhydrous K₂CO₃, methallyl chloride (2.77 g, 30.5 mmoL), Nal(1.07 g, 6.7 mmoL). The resulting slurry was stirred at ambienttemperature under N₂ for 24 h. 50 mL of ice-cold water was addedfollowed by 2N KHSO₄ solution until pH was 1. EtOAc (100 mL) was addedand the mixture was stirred for 0.75 h. Combined organic layer wascollected and washed with brine, dried over MgSO₄, and evaporated toyield the desired product, 23.04.

The product 23.04 from the preceding step (2.7 g, 8.16 mmoL) wasdissolved in dioxane (20 mL) and treated with freshly prepared 1N LiOH(9 mL). The reaction mixture was stirred at ambient temperature under N₂for 20 h. The reaction mixture was taken in EtOAc and washed with H₂O.The combined aqueous phase was cooled to 0° C. and acidified to pH 1.65using 1N HCl. The turbid mixture was extracted with EtOAc (2×100 mL).Combined organic layer was washed with brine, dried over MgSO₄, andconcentrated to give the desired acid, 23.05 (3.40 g).

To a suspension of NaBH(OAc)₃ (3.93 g, 18.5 mmoL) in CH₂Cl₂ (55 mL) wasadded a solution of product 23.05 from preceding step in anhydrousCH₂Cl₂ (20 mL) and acetic acid (2 mL). The slurry was stirred at ambienttemperature for 20 h. Ice cold water (100 mL) was added to the slurryand stirred for ½ hr. Organic layer was separated, filtered, dried andevaporated to yield the desired product, 23.06.

To a solution of the product 23.06 from preceding step (1.9 g) in MeOH(40 mL) was treated with excess of CH₂N₂/Et₂O solution and stirred forovernight. The reaction mixture was concentrated to dryness to yield acrude residue. The residue was chromatographed on silica gel, elutingwith a gradient of EtOAc/hexane to afford 1.07 g of the pure desiredproduct, 23.07.

To a solution of product 23.07 from preceding step (1.36 g) in anhydrousCH₂Cl₂ (40 mL) was treated with BF₃. Me₂O (0.7 mL). The reaction mixturewas stirred at ambient temperature for 20 h and quenched with sat.NaHCO₃ (30 mL) ad stirred for ½ hr. Organic layer was separated andcombined organic layer was washed with brine, dried over MgSO₄,concentrated to give crude residue. The residue was chromatographed onsilica gel eluting with a gradient of EtOAc/hexane to afford 0.88 g ofthe desired compound, 23.08.

To a solution of the product 23.08 (0.92 g) from preceding step in MeOH(30 mL) was added 10% Pd/C (0.16 g) at room temperature and hydrogenatedat ambient temperature under 1 atm. Pressure. The reaction mixture wasstirred for 4 h and concentrated to dryness to yield the desiredcompound, 23.01.

Preparation of Intermediate 50.01

To a solution of 50.02 (15 g) in MeOH (150 mL) was added conc HCl (3–4mL) and the mixture was refluxed for 16 h. The reaction mixture wascooled to room temperature and concentrated. The residue was taken indiethyl ether (250 mL) and washed with cold saturated sodium bicarbonatesolution, and brine. The organic layer was dried (Na₂SO₄) andconcentrated to afford the methyl ester 50.03 (12.98 g) which wascarried forward without further purification.

The methyl ester 50.03 from above was dissolved in methylene chloride(100 mL) and cooled to −78° C., under nitrogen atmosphere. DIBAL (1.0 Msolution in methylene chloride, 200 mL) was added dropwise over 2 hperiod. The reaction mixture was warmed to room temperature over 16 h.The reaction mixture was cooled to 0° C. and MeOH (5–8 mL) was addeddropwise. A solution of aqueous 10% sodium potassium tartarate (200 mL)was slowly added with stirring. Diluted with methylene chloride (100 mL)and separated the organic layer (along with some white precipitate). Theorganic layer was washed with 1 N HCl (250 mL), brine (200 mL), dried(Na₂SO₄) and concentrated to provide the alcohol 50.04 (11.00 g) as aclear oil.

The alcohol 50.04 from above was dissolved in methylene chloride (400mL) and cooled to 0° C. under nitrogen atmosphere. PCC (22.2 g) wasadded in portions and the reaction mixture was slowly warmed to roomtemperature over 16 h. The reaction mixture was diluted with diethylether (500 mL) and filtered through a pad of celite. The filtrate wasconcentrated and the residue was taken in diethyl ether (500 mL). Thiswas passed through a pad of silica gel and the filtrate was concentratedto provide the aldehyde 50.05 which was carried forward without furtherpurification.

The aldehyde 50.05 from above was converted to the desired material50.01 using essentially the method of Chakraborty et al (Tetrahedron,1995, 51(33), 9179–90).

Preparation of Intermediate 51.01

The required intermediate 51.01 was obtained from the aldehyde 51.02using the literature described procedure (T. K. Chakraborty et al.,Tetrahedron, 1995, 51 (33), 9179–90).

Preparation of Compound 5000:

To a stirred solution of N-Boc-(S)-valinol 5000a (10.0 g, 49.2 mmol),phthalimide (7.40 g, 50.3 mmol) and triphenylphosphine (13.0 g, 49.6mmol) in anhydrous THF (100 mL) at 0° C. was addeddiisopropylazodicarboxylate (DIAD, 9.8 mL, 49.4 mmol). The resultingsolution was then stirred at rt for 18 h before it was concentrated todryness. The residue was dissolved in CH₂Cl₂ and purified by silica gelflash chromatography (10–40% EtOAc in hexanes) to give product 5000b.

To a stirred solution of 5000b (8.8 g, 26.5 mmol) in methanol (100 mL)at rt was added hydrazine monohydrate (1.4 mL, 28.8 mmol) and theresulting solution was stirred at rt for 18 h. Additional hydrazinemonohydrate (0.5 mL, 10.3 mmol) was added and the mixture was brought toreflux and stirred for 4 h before it was cooled to rt. The precipitatewas filtered off and the solution was concentrated to dryness. Theresidue was dissolved in CH₂Cl₂ and the precipitate was again filteredoff. After concentration, a yellow oil was obtained (8.0 g, quant.).

To a solution of 5000c (1.0 g, 4.94 mmol) in CH₂Cl₂ I (100 mL) at −30°C. in an acetone bath was added 3-chloropropyl sulfonyl chloride (0.60mL, 4.93 mmol) and triethylamine (1.10 mL, 7.89 mmol). The resultingsolution was warmed to rt along with the bath and stirred for 18 h.Additional CH₂Cl₂ and 1 N Na₂CO₃ solution were added and the layers wereseparated. The aqueous solution was extracted with CH₂Cl₂ (2×100 mL).The organic solutions were combined, filtered, dried (MgSO₄) andconcentrated. The residue was purified by flash column chromatographyusing 10–40% acetone/hexanes to afford 1.0 g (59%) of 5000d.

The suspension of 5000d (1.0 g, 2.92 mmol) and sodium hydride (0.32 g,60%, 8.0 mmol) in anhydrous DMF (100 mL) was stirred at rt for 8 h.After cooled to 0° C., 5% aqueous phosphoric acid solution (110 mL) wascautiously added followed by EtOAc (150 mL). The layers were separatedand the organic solution was washed with 5% aqueous phosphoric acidsolution (100 mL) and saturated sodium bicarbonate solution (2×100 mL)before it was dried, filtered and concentrated. The residue was purifiedby silica gel flash chromatography (0–40% acetone in hexanes) to give0.63 g product 5000e (70%).

The solution of 5000e (0.62 g, 2.02 mmol) in 4 N HCl in dioxane wasstirred at rt for 4 h. It was then concentrated to dryness in vacuo togive 0.60 g product 5000 (quant.).

Preparation of Compound of Formula 5001

Compound 5001a was prepared from 5000c and 4-bromobutyryl chlorideaccording to the procedures described for the preparation of compound5000d.

Preparation of Compound of Formula 5002

Compound 5002a was prepared from 5000c and 2-bromoethyl chloroformateaccording to the procedures described for the preparation of compound5000d.

Compound 5002b was prepared from 5002a according to the proceduresdescribed for the preparation of compound 5000e.

Compound 5002 was prepared from 5002b according to the proceduresdescribed for the preparation of compound 5000 (Step 5).

Preparation of Compound of Formula 5003

Compound 5003b was prepared from N-Boc-(S)-tert-leucinol 5003a accordingto the procedures described for the preparation of compound 5000b (Step1).

Compound 5003 was prepared from 5003b according to the proceduresdescribed for the preparation of compound 5000 (Step 5).

Preparation of Compound of Formula 5004

Compound 5004a was prepared from 5003b according to the proceduresdescribed for the preparation of compound 5000c (Step 2).

Compound 5004b was prepared from 5004a according to the proceduresdescribed for the preparation of compound 5000d (Step 3).

Compound 5004c was prepared from 5004b according to the proceduresdescribed for the preparation of compound 5000e (Step 4).

Compound 5004 was prepared from 5004c according to the proceduresdescribed for the preparation of compound 5000 (Step 5).

Preparation of Compound of Formula 5005

Compound 5005a was prepared from 5004a according to the proceduresdescribed for the preparation of compound 5002a (Step 1).

Compound 5005b was prepared from 5005a according to the proceduresdescribed for the preparation of compound 5002b (Step 2).

Compound 5005 was prepared from 5005b according to the proceduresdescribed for the preparation of compound 5000 (Step 5).

Preparation of Compound of Formula 5006

Compound 5006a was prepared from 5004a and 4-bromobutyryl chlorideaccording to the procedures described for the preparation of compound5000d.

Compound 5006 was prepared from 5006a according to the proceduresdescribed for the preparation of compound 5000 (Step 5).

Preparation of Compound of Formula 5007

Compound 5007a was prepared from 5004a and2-carbomethoxy-3-thiophenesulfonyl chloride according to the proceduresdescribed for the preparation of compound 5000d.

To the solution of ester 5007a (4.65 g, 11.1 mmol) in anhydrous toluene(40 mL) at −78° C. was added a solution of DIBAL-H in toluene (23.0 mL,34.5 mmol). The mixture was stirred at −78° C. for 20 min and at rt for2 h. Methanol (20 mL) was added followed by 10% aqueous citric acidsolution (100 mL). After stirred for 5 min, EtOAc (200 mL) was added andlayers were separated. The aqueous solution was extracted with EtOAc(2×100 mL). The organic solutions were combined, dried (MgSO₄), filteredand concentrated. The residue was purified by flash columnchromatography using 10–50% acetone/hexanes to give 4.6 g (quant.) of5007b.

To a solution of 5007b (1.04 g, 2.65 mmol) in CH₂Cl₂I (50 mL) at −0° C.was added methanesulfonyl chloride (0.23 mL, 2.97 mmol) andtriethylamine (0.80 mL, 5.74 mmol). The mixture was warmed to rt alongwith ice bath and stirred for 18 h. EtOAc (200 mL) and 5% H₃PO₄ solution(100 mL) was added and the layers were separated. The organic solutionswere washed with 1 N sodium carbonate solution (100 mL) before it wasdried (MgSO₄), filtered and concentrated. The residue was purified byflash column chromatography using 10–50% acetone/hexanes to give 0.80 g(73%) of 5007c.

The suspension of 5007d (1.17 g, 2.85 mmol) and cesium carbonate (1.40g, 4.30 mmol) in anhydrous DMF (100 mL) was stirred at rt for 18 h.Water (50 mL), brine (50 mL) and EtOAc (300 mL) were added and thelayers were separated. The organic solution was washed water (3×150 mL)before it was dried, filtered and concentrated to give 0.99 g of thedesired product 5007d (93%).

Compound 5007 was prepared from 5007d according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5008

Compound 5008b was prepared from 5003a and 5008a according to theprocedures described for the preparation of compound 5000b.

Compound 5008 was prepared from 5008b according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5009

To a solution of 5008b (1.02 g, 2.93 mmol) in CH₂Cl₂ (50 mL) at −18° C.was added m-chloroperoxybenzoic acid (3.03 g, 17.6 mmol) and theresulting solution was stirred at −18° C. for 1 h before it was placedin a refrigerator overnight (16 h). After stirred at rt for another 6 h,additional CH₂Cl₂ was added and the solution was washed with 10% NaHSO₄and 1 N Na₂CO₃. The organic solutions was dried (MgSO₄), filtered andconcentrated. The residue was purified by flash column chromatographyusing 5–60% acetone/hexanes to afford 0.49 g (46%) of product 5009a.

Compound 5009 was prepared from 5009a according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5010

Compound 5010a was prepared from 5004a and methyl2-(chlorosulfonyl)benzoate according to the procedures described for thepreparation of compound 5000d.

Compound 5010b was prepared from 5010a according to the proceduresdescribed for the preparation of compound 5007b.

Compound 5010c was prepared from 5010b according to the proceduresdescribed for the preparation of compound 5007c.

Compound 5010d was prepared from 5010c according to the proceduresdescribed for the preparation of compound 5007d.

Compound 5010 was prepared from 5010d according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5011

The suspension of 5010a (0.60 g, 1.45 mmol) and cesium carbonate (0.707g, 2.17 mmol) in anhydrous DMF was stirred at 40° C. for 18 h. Water (50mL), brine (50 mL) and EtOAc (150 mL) were added and the layers wereseparated. The organic solution was washed water (3×80 mL) before it wasdried, filtered and concentrated to give 0.17 g of the desired product5011a (31%).

Compound 5011 was prepared from 5011a according to the proceduresdescribed in Step 5 of the preparation of compound 5000.

Preparation of Compound of Formula 5012

Compound 5012a was prepared from 5003a and glutarimide according to theprocedures described for the preparation of compound 5000b.

Compound 5012 was prepared from 5012a according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5013

The solution of amine 5004a (3.0 g, 13.9 mmol) and 1,4-butane sultone(1.8 mL, 17.7 mmol) in anhydrous THF (25 mL) was refluxed for 16 h. Moresultone (0.6 mL, 5.89 mmol) was added and the mixture was refluxed foranother 4 h. Phosphorus oxychloride (2.6 mL, 27.9 mmol) was added andthe solution was stirred at rt for 4 h. After cooled to 0° C., 50% w/wNaOH solution was added slowly along with water (30 mL) until PH isgreater than 12. Ether (200 mL) was then added and layers wereseparated. The aqueous solution was extracted with THF/diethyl ether(1:1, 150 mL) twice. Organic solutions were combined, dried (MgSO₄),filtered and concentrated. The residue was purified by flash columnchromatography using 10–50% acetone/hexanes to give 1.49 g of 5013a(32%).

Compound 5013 was prepared from 5013a according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5014

To the suspension of 5014a (10.0 g, 41.1 mmol), HOOBt (8.7 g, 53.3mmol), EDCI (10.0 g, 52.2 mmol) and ammonium chloride (8.90 g, 166 mmol)in anhydrous DMF (400 mL) at rt was added 4-methylmorpholine (22.5 mL,204.5 mmol). The mixture was stirred at rt for 70 h. Brine (150 mL) and5% aqueous phosphoric acid solution (150 mL) were added followed byEtOAc (800 mL). The layers were separated and the organic solution waswashed with 5% aqueous phosphoric acid solution (400 mL) and saturatedsodium bicarbonate solution (2×400 mL) before it was dried, filtered andconcentrated to give 8.35 g product 5014b (84%).

The solution of amide 5014b (8.35 g, 34.5 mmol) in anhydrous THF (100mL) at rt was added a solution of borane methyl sulfide complex intoluene (43.0 mL, 86.0 mmol) and the mixture was refluxed for 4 h. MoreTHF (100 mL) was added and 3 N HCl solution was added slowly until nogas evolution was observed. To the mixture was added 50% w/w NaOHsolution slowly until PH is greater than 12. Ether (200 mL) was thenadded and layers were separated. The aqueous solution was extracted withTHF/diethyl ether (1:1, 150 mL) twice. Organic solutions were combined,dried (MgSO₄), filtered and concentrated to give 6.50 g of the product5014c (83%).

To the solution of the amine 5014c (0.80 g, 3.50 mmol) and N-carbethoxyphthalimide 5014d (0.90 g, 4.11 mmol) in anhydrous THF (50 mL) at rt wasadded triethylamine (1.0 mL, 7.17 mmol). The mixture was stirred at rtfor 18 h. EtOAc (100 mL) and 5% aqueous phosphoric acid solution (100mL) were added and the layers were separated. The organic solution waswashed with 5% aqueous phosphoric acid solution (80 mL) and dried,filtered and concentrated. The residue was purified by silica gel flashchromatography (5–50% EtOAc in hexanes) to give 0.82 g product 5014e(65%).

Compound 5014 was prepared from 5014e according to the proceduresdescribed for the preparation of compound 5000.

Preparation of Compound of Formula 5015

Compound 5015 was prepared from 5014c according to the procedures steps1–5 described for the preparation of compound 5007.

Preparation of Compound of Formula 5016

Compound 5016b was prepared from 5016a according to the procedure step 1described for the preparation of compound 5014b.

Compound 5016c was prepared from 5016b according to the procedure step 2described for the preparation of compound 5014c.

Compound 5016 was prepared from 5016c according to the procedure steps3–7 described for the preparation of compound 5010.

Preparation of Compound of Formula 5017

Compound 5017 was prepared from 5016c and 1,4-butane sultone accordingto the procedures in steps 1–2 described for the preparation of compound5013.

Preparation of Compound of Formula 5018

Compound 5018 was prepared from 5003a and morpholine 3,5-dione accordingto the procedures in steps 1–2 described for the preparation of compound5012.

Preparation of Compound of Formula 5019

Compound 5019 was prepared from 5003a and 3,3-dimethyl glutarimideaccording to the procedures in steps 1–2 described for the preparationof compound 5012.

Preparation of Compound of Formula 5020

To the solution of chlorosulfonyl isocyanate (0.80 mL, 9.25 mmol) inanhydrous CH₂Cl₂ (20 mL) at 0° C. was added slowly benzyl alcohol (0.96mL, 9.25 mmol). The resulting solution was stirred at 0° C. for 30 minbefore it was added slowly to a solution of amine 5020a (2.0 g, 9.25mmol) in anhydrous CH₂Cl₂ at 0° C. The mixture was stirred at 0° C. for1 h and at rt for another hour before it was concentrated to dryness.The residue was dissolved in EtOAc and washed with 1 N HCl solutiontwice and brine once. It was then dried, filtered and concentrated. Theproducts were purified by silica gel flash chromatography (20–70%acetone in hexanes) to give product 5020b (3.11 g, 78%).

To the solution of 5020b (1.60 g, 3.73 mmol), triphenylphosphine (1.46g, 5.57 mmol) and 3-bromo-1-propanol (0.36 mL, 4.12 mmol) in anhydrousCH₂Cl₂ (40 mL) at 0° C. was added diisopropylazodicarboxylate (DIAD,1.10 mL, 5.55 mmol). The resulting solution was then stirred at rt for 2h before it was concentrated in vacuo to dryness. The residue waspurified by silica gel flash chromatography (10–40% acetone in hexanes)to give the product 5020c (1.62 g, 79%).

The suspension of 5020d (1.61 g, 2.93 mmol) and cesium carbonate (1.43g, 4.39 mmol) in anhydrous DMF (100 mL) was stirred at rt for 18 h.Water (50 mL), brine (50 mL) and EtOAc (300 mL) were added and thelayers were separated. The organic solution was washed water (3×150 mL)before it was dried, filtered and concentrated to give the desiredproduct 5020d (1.40 g, quant.).

The mixture of 5020d (1.40 g, 2.98 mmol) and 10% Pd—C (wet basis) inabsolute ethanol (50 mL) and methanol (50 mL) was vigorously stirred atrt for 2.5 h. The catalyst was filtered off through a celite pad to giveproduct 5020e (1.10 g, quant.).

The solution of 5020e in 4 N HCl in dioxane was stirred at rt for 4 h.It was then concentrated to dryness in vacuo to give product 5020.

Preparation of Compound of Formula 5021

Compound 5021 was prepared from 5003a and tetramethylene glutarimideaccording to the procedures in steps 1–2 described for the preparationof compound 5012.

Preparation of Compound of Formula 5022

The mixture of anhydride 5022a and amine 5004a in anhydrous toluene wasbrought to reflux and stirred for 46 h before it was cooled andconcentrated in vacuo. The residue was purified by silica gel flashchromatography (5–40% EtOAc in hexanes) to give the product 5022b (2.90g, 79%).

The compound 5022b was treated with 4 N HCl for 30 min at rt andconcentrated in vacuo to give product 5022.

PREPARATION OF EXAMPLES

Preparation of Compound of 5146

To the solution of 5020b (2.0 g, 4.66 mmol), triphenylphosphine (1.83 g,6.99 mmol) and 2-bromo-ethanol (0.31 mL, 5.12 mmol) in anhydrous CH₂Cl₂(30 mL) at 0° C. was added diisopropylazodicarboxylate (DIAD, 0.996 mL,6.99 mmol). The resulting solution was then stirred at rt for 2 h beforeit was concentrated in vacuo to dryness. The residue was purified bysilica gel flash chromatography (10–40% acetone in hexanes) to give theproduct 5051a (1.50 g, 63%).

The suspension of 5051a (1.5 g, 2.79 mmol) and cesium carbonate (1.36 g,4.19 mmol) in anhydrous DMF (100 mL) was stirred at rt for 18 h. Water(50 mL), brine (50 mL) and EtOAc (300 mL) were added and the layers wereseparated. The organic solution was washed water (3×150 mL) before itwas dried, filtered and concentrated to afford 1.37 g of crude product.Purified via flash column (10–30% Acetone-hexane) to afford 0.98 g of5051b (82%).

To compound 5051b (0.98 g, 2.15 mmol, 1 equiv.) was added 4 M HCl indioxane (25 mL) at room temp. Stirred for 1 hr. TLC showed no startingmaterial. Evaporated off the solvent and azeotroped with hexane and thenwith ether. Washed out the non-polar material with ether and kept underhigh vac. over the weekend to give the product as a pale yellow solid(842 mg, quant.). Product was used without purification.

To the amine hydrochloride 1.04 (3 g, 9.4 mmol) in Dichloromethane (50ml) was added 50 ml of saturated NaHCO₃. Stirred vigorously at icetemperature for 5 min. Stopped stirring and phosgene (2 equiv. 20% intoluene, 10 mL) was syringed out to the lower layer and restored thevigorous stirring immediately. Checked the TLC at times and after 2 hrsit showed complete consumption of starting material and then separatedthe layers. Washed the water layer one more time with dichloromethane (3ml) and dried over anhydrous sodium sulfate. Filtered and evaporated offthe solvent using rotary evaporator under reduced pressure without hotbath to half the volume and then flushed N₂ for 15 minutes. Diluted to33.5 mL with dichloromethane and used as 0.28 M solution for furthercouplings.

To the amine 5051c, prepared as described before (741 mg, 2.09 mmol, 1equiv.) in DCM (10 ml) was added DIPEA (8 equiv., 2.19 mL, 12.54 mmol)at ice temperature. Added isocyanate 5051d (1 equiv, 7.46 of 0.028Msolun) under N₂ atm and stirred for 30 min at ice temperature and 90 minat room temperature. Quenched with 10% citric acid and extracted withEtOAc and washed with brine. Dried over anhydrous sodium sulfate andfiltered and evaporated off the solvent. The crude product was purifiedvia flash column (10–40% % Acetone-hexane) to afford 800 mg of 5051e asa white solid (58%). ¹H NMR (CDCl₃, 300 MHz), δ, 7.4(m, 5H), 5.3(bs,2H), 4.4(d, 2H,), 4–3.6 (m, 6H), 3.6 (s, 3H), 3.25 (m, 2H), 3(m, 2H),1.02–0.98 (m, 25H).

The mixture of 5051e (0.600 g, 0.904 mmol) and 10% Pd—C (10% wt., 60 mg)in methanol (10 mL) was vigorously stirred at rt for 1.5 h. The reactionmixture was filtered through a celite pad to give product 5051f (0.45 g,94%.).

To the sulfamide 5051f (400 mg, 0.756 mmol, 1 equiv.) in DMF (10 mL) atice temperature, added Cs₂CO₃ (368 mg, 1.5 equiv, 1.134 mmol) and Mel(3.78 mmol, 5 equiv., 0.355 mL) under nitrogen atmosphere. Stirred atroom temperature for overnight. As the TLC and LCMS showed no startingmaterial, quenched with water and extracted with EtOAC. Washed 4 timeswith water and with brine and dried over anhydrous sodium sulfate.Filtered and evaporated off the solvent and purified via flash column(20–40% acetone-hexane) to afford 390 mg of 5051g (95%). ¹H NMR (CDCl₃,300 MHz) 4.4(d, 1H,), 3.99–4.01 (d, 1H), 3.8 (d, 2H), 3.7(s, 3H), 3.6(m, 2H), 3.25(m, 2H), 3.01 (m, 3H), 2.8(s, 3H), 1.4(m, 1H), 1.2(m, 1H),1.00–0.98 (m, 24H).

To the methyl ester, 5051g (300 mg, 0.607 mmol, 1 equiv.) in dioxane (10ml) was added LiOH (1.8 mL, 1N in water, 3 equiv) and stirred overnight.Quenched with 1 N HCl and extracted with EtOAC. Washed with brine anddried over anhydrous sodium sulfate. Filtered and evaporated off thesolvent to give the crude product (290 mg, 90%).

To solution of the amine 10.11, prepared as described before (16.51 mg,0.079 mmol, 1.2 equiv.), and 5051h (35 mg, 0.066 mmol, 1 equiv.) in DMF(10 ml) at 0° C. was added HATU (1.2 equiv., 0.079 mmol, 30.18 mg)followed by DIPEA (8 equiv., 92.44 □L, 0.529 mmol). Stirred for 1 h atice temperature and then 2 h at room temperature. Quenched with 1 N HCland extracted with EtOAC. Washed with sat'd sodium bicarbonate and thenwith brine. Dried over anhydrous sodium sulfate, filtered and evaporatedoff the solvent to give the product (52 mg, 100%).

To the hydroxy amide 5051i (60 mg, 0.087 mmol, 1 equiv.) in 1:1 mixtureof DMF/toluene (6 mL) at ice temperature was added EDCI.HCl (167 mg, 10equiv., 0.878 mmol) and dichloroacetic acid (36.29 □L, 5 equiv., 0.439mmol) and stirred for 5 min. Then stirred at room temperature forfurther 3 hrs. Quenched with brine and washed with 1 N HCl followed bysat'd NaHCO₃ and again with brine. Dried over anhydrous sodium sulfate,filtered and evaporated off the solvent. The crude product was purifiedby preparative TLC (40% acetone-hexane) to afford 25.2 mg of 5146 (43%).LRMS, m/z, 682[(M+1)], 375.

Preparation of Compound of Formula 5237

To compound 5051b (1.16 g, 2.5 mmol) in methanol was added Pd/C (5% bywt, 116 mg) under N₂ atmosphere after evacuation. Evacuated again andstirred under H₂ for 90 min. TLC showed complete consumption of startingmaterial. Filtered and evaporated off the solvent to afford 5052a (819mg, 100%).

To the amino compound (285 mg, 1 equiv.) in DMF (10 ml) added 2-iodopropane (5 equiv.) and Cesium carbonate (1.5 equiv.) at ice temperatureand stirred for overnight. Temperature of the reaction mixture wasslowly raised to room temperature. Quenched with water and extractedwith EtOAc and washed the combined organic extracts with brine. Driedover anhydrous sodium sulfate, filtered and evaporated off the solvent.The crude product, 5052b was used as it is for next step (310 mg, 96%).¹H NMR (CDCl₃, 300 MHz) 4.5(d, 1H,), 3.8–3.6 (m, 2H), 3.4–3.2 (m, 2H),3.01–2.8(m, 2H), 1.9 (m, 1H), 1.4 (s, 9H), 1.2(dd, 6H), 0.98(s, 9H).

Compound 5052b was dissolved in 4 N HCl in dioxane at rt and thesolution was stirred for 1 hr. TLC showed no starting material.Evaporated off the solvent and azeotroped with hexane and then withether. Kept under high vac. for overnight to afford 221 mg of 5052c(96%).

Dissolved the amine salt (180 mg, 0.60 mmol, 1 equiv.) in DCM (5 ml) andadded 5 mL of NaHCO₃ (sat'd) at ice temperature. Stirred vigorously for2 min. Stopped stirring and syringed out phosgene (2 equiv.) to thereaction mixture and restored the vigorous stirring. After 90 min.separated the layers and dried over anhydrous sodium sulfate. Filteredand evaporated off the solvent without hot bath under vac. Diluted withDCM and kept as stock solution of 0.02 M.

To a mixture of acid 1.17 (500 mg, 1.37 mmol, 1 equiv.) and aminehydrochloride (317.8 mg, 1.37 mmol, 1 equiv.) in DMF at ice temperaturewas added HATU (1.2 equiv. 619 mg) and DIPEA (6 equiv., 8.15 mmol, 1.42mL) under N₂ and stirred for overnight. The temperature was slowlyallowed to raise to room temperature. Quenched with 1 N HCl andextracted with EtOAc. Washed with NaHCO₃ (sat) and then with brine.Washed with ice-cold water (5×20 ml) and again with brine. Dried overanhydrous Na₂SO₄. Filtered and evaporated off the solvent to afford 580mg of 5052e (77%).

To the crude hydroxy amide 5052e (1.05 mmol, 580 mg, 1 equiv.) in DCM(15 mL) at room temperature, was added Dess-Martin Periodinane (897 mg,2.11 mmol, 2 equiv.). Stirred for 5 h at rt. Quenched with saturatedNaHCO₃ and sodium bisulfite and extracted with EtOAc. Washed with brineand dried over anhydrous sodium sulfate. Filtered and evaporated off thesolvent. Crude product was purified by flash column (10–40%acetone-hexane) to afford 450 mg of the ketoamide product. The productwas dissolved in 4 N HCl solution in dioxane and stirred at rt for 3 hbefore it was 10 concentrated to dryness in vacuo to give 5052f (0.40 g,77%).

To the amine salt, 5052f, (20 mg, 0.041 mmol, 1 equiv.) in DCM (5 ml)was added DIPEA (6 equiv.) at ice temperature. Added isocyanate, 5052d(1.1 equiv, 0.045 mmol, 2.27 mL of 0.02M solun) under N2 atm and stirredfor 30 min at ice temperature and 90 min at room temperature. Quenchedwith citric acid and extracted with EtOAc and washed with brine. Driedover anhydrous sodium sulfate and filtered and evaporated off thesolvent. The crude product was purified via flash column (10–40%acetone-hexane) to afford 12 mg of 5237 (40%).

Preparation of Compound 5250

Compound 5250a was prepared from compound 20.03 according to theprocedures described for the preparation of 20.08. To the solution ofamine 5010 (0.817 g, 2.68 mmol) and carbamate 5250a (0.976 g, 2.06 mmol)in anhydrous DCM (60 mL) at 0° C. was added DIPEA (0.90 mL, 5.15 mmol).The solution was allowed to warm to rt along with ice bath and stirredfor 18 h before it was concentrated. The residue was dissolved in EtOAcand washed with 5% H₃PO₄ solution and saturated sodium bicarbonatesolution. It The products were purified by silica gel flashchromatography to give product 5250b (1.07 g, 86%).

The solution of methyl ester 5250b (1.06 g, 1.76 mmol) and LiOH (0.105g, 4.40 mmol) in THF/MeOH/H₂O (1:1:1, 30 mL) was stirred at roomtemperature for 4 h. Methanol and THF were removed under reducedpressure. The aqueous solution was acidified to PH˜2 using 1 N aqueousHCl solution (50 mL) and saturated with solid sodium chloride before itwas extracted with EtOAc (3×150 mL). The organic solutions werecombined, dried (MgSO₄), filtered and concentrated in vacuo to give awhite solid 5250c (quantitative).

To the suspension of 5250c (0.052 g, 0.088 mmol), HOOBt (0.022 g, 0.132mmol), EDCI (0.027 g, 0.141 mmol) and amine hydrochloride 13.01 (0.030g, 0.132 mmol) in anhydrous DMF (6 mL) and DCM (6 mL) at −20° C. wasadded 4-methylmorpholine (0.030 mL, 0.273 mmol). The mixture was stirredat −20° C. for 20 min and then in a refrigerator for 18 h. Brine (30 mL)and 5% aqueous phosphoric acid solution (30 mL) were added followed byEtOAc (100 mL). The layers were separated and the organic solution waswashed with 5% aqueous phosphoric acid solution (50 mL) and saturatedsodium bicarbonate solution (2×50 mL) before it was dried, filtered andconcentrated to give product 5250d (quantitative).

The mixture of hydroxyamide 5250d and Dess-Martin periodinane (g) inCH₂Cl₂ was stirred for 2 h, quenched with saturated Na₂S₂O₃ solution andsaturated NaHCO₃ solution. After layers were separated, the organicsolution was extracted with DCM twice and the combined organic solutionwas dried, filtered and concentrated to give g product 5250 (0.048 g,72%, two steps).

Preparation of Compound 5648

To a cooled solution (0° C.) of (S)-tert-leucinol (5.0 g, 42.7 mmol) inCH₂Cl₂ (100.0 mL) was added benzyl chloroformate (6.7 mL, 47.0 mmol),followed by Hunig's base (9.3 mL, 53.3 mmol). The reaction mixture wasallowed to warm to room temperature overnight, diluted with ethylacetate (500 mL), washed with 10% KH₂PO₄, followed by saturated NaHCO₃and brine. The organic layer was dried over MgSO₄ and concentrated toyield 5500a (10.7 g, 100%).

To a cooled solution (0° C.) of 5500a (10.7 g, 42.7 mmol) in CH₂Cl₂(100.0 mL) was added pyridine (20.0 mL), followed by methanesulfonylchloride (3.63 mL, 47.0 mmol). The reaction mixture was allowed to warmto room temperature overnight, concentrated, redissolved in ethylacetate (500 mL), washed with saturated NaHCO₃ and brine. The organiclayer was dried over MgSO₄, concentrated and purified by flashchromatography over SiO₂ using ethyl acetate/hexane (1:4) to yield 5500b(14.0 g, 100%).

To a solution of 5500b (3.1 g, 9.9 mmol) in PhMe (72 mL) containingwater (400 μL) was added TBAB (582 mg, 1.8 mmol), K₂CO₃ (2.72 g, 1.97mmol), 1-hydroxypyridine (937 mg, 9.85 mmol). The reaction mixture wasrefluxed overnight with stirring, filtered, evaporated and concentrated.The crude was purified by flash chromatography over SiO₂ using ethylacetate/CH₂Cl₂ (1:9 to 1:1) to yield 5500c (1.15 g, 35%).

To a solution of 5500c (1.15 g) in MeOH (50 mL) was added Pd/C(10% w/w,450 mg) and placed in a Parr shaker under a hydrogen atmosphere (40PSI). 5500d was formed quantitatively when the reaction was stoppedafter 1.2 h and 5500e was formed quantitatively after 4 h. In eithercase the reaction mixture was filtered over a short pad of celite toyield the corresponding amine.

Saturated NaHCO₃ (7.0 mL) was added to an ice-cold solution of 5500d(194.0 mg, 1 mmol) in CH₂Cl₂ (7 mL). The reaction mixture was stirredvigorously for 10 min. and COCl₂ (1.85 M solution in toluene, 1.35 mL)was added to it and stirring was continued at room temperature for 1 h.The organic layer was dried over MgSO₄, filtered and concentrated tohalf the volume to yield 5500f as a solution in CH₂Cl₂. 5500f was storedas a 0.05 M solution in CH₂Cl₂.

Compound 5500g was synthesized using the procedure of step 5 in thepreparation of 5500.

To a cooled solution (0° C.) of the acid (1.17, 368.5 mg) and (10.11,565.3 mg) in DMF (10.0 mL) was added HATU (1.03 g), followed by DIPEA1.382 mL). The reaction mixture was stirred at 0° C. for 1 h and at roomtemperature for 2 h, diluted with ethyl acetate (20.0 mL), washed with1N HCl, brine, dried over NaHCO₃, filtered, concentrated.

To the crude in PhMe/DMSO (10.0 mL, 1:1) at 0° C. was added EDCI 5.2 g),followed by dichloroacetic acid (447 μL). The ice bath was removed andthe reaction was stirred at room temperature for 2 h. To it was addedEtOAc (75 mL) the reaction mixture was washed with H₂O (25.0 mL), withsaturated NaHCO₃ and brine, then purified over SiO₂ using acetone/hexane(1:9 to 9:1) to yield 5500h.

Compound 5500h was dissolved in 4N HCl in dioxane (25 mL). The reactionwas stirred at room temperature for 30 min. and concentrated to yield awhite solid, 5500i (350.0 mg).

To a cooled solution (0° C.) of the amine hydrochloride 5500i (25.0 mg,0.051 mmol) in CH₂Cl₂ (2.0 mL) was added 5500f (2.5 mL, 0.135 mmol),followed by DIPEA (68 uL, 0.4 mmol). The reaction mixture was stirred atroom temperature for 1.2 h, diluted with ethyl acetate (20.0 mL), washedwith 3% citric acid, brine, dried over NaHCO₃, filtered, concentratedand purified over SiO₂ using acetone-hexane (1:9) to yield 5648 (10.0mg). LCMS 641.2 (M+H).

Preparation of Compound 5644

Compound 5644 was synthesized using the procedures described for thepreparation of 5648. LCMS 645.0 (M+H).

A number of analogs of 5644, described in Table 2 were prepared from5500g using the procedures described in the preparation of 5644.

Preparation of Compound 5632

Compound 5632a was prepared from (S)-N-boc valinol according to theprocedures in the preparation of 5500g (steps 1–6). Compound 5632 wassynthesized according to procedures described for the preparation of5648. LCMS: 631.1 (M+H)

A number of analogs of compound 5632 in Table 2 were prepared from 5632ausing the same procedures as in the preparation of 5632.

Preparation of Compound 5665

To a cooled (0° C.) of amine 5004a (560.0 mg, 2.58 mmol) in CH₂Cl₂ (15.0mL) was added chloropropyl isocyanate (Aldrich, 531 mL, 5.16 mmol) andthe reaction mixture was stirred at room temperature for 12 h, washedwith saturated NaHCO₃, brine, dried over MgSO₄, filtered, concentratedand purified over SiO₂ using ethyl-acetate-hexane (1:1) to yield 5520a(660 mg, 1.91 mmol, 76%).

To a cooled solution of 5520a (660.0 mg, 1.96 mmol) in THF (30 mL) wasadded NaH. (60% dispersion in mineral oil, 313.0 mg, 7.84 mmol). Thereaction mixture was allowed to warm up to room temperature over 4 hour,carefully quenched with ice cold water, extracted with CH₂Cl₂. Theorganic layer was washed with brine, dried over MgSO₄ filtered andconcentrated. The crude was purified over SiO₂ using ethylacetate-hexane (1:1) to yield 5520b (220.0 mg, 1.0 mmol).

To 5520b (660.0 mg, 1.96 mmol) was added 4N HCl in dioxane (25 mL). Thereaction was stirred at room temperature for 30 min and concentrated toyield a white solid which was dissolved in CH₂Cl₂ (7.0 mL) and sat'dNaHCO₃ (7.0 mL) was added to it. After stirring vigorously for 10 min,the layers were allowed to separate and phosgene (2.5 equiv.) was addedto the organic layer in one shot. Vigorous stirring was resumedimmediately, the ice bath was removed and stirring was continued for 1h, the layers were separated. The organic layer was dried over MgSO₄,filtered and concentrated to half to its volume and 5520c was stored asa 0.05 M solution in CH₂Cl₂.

To a cooled solution (0° C.) of the amine hydrochloride 5500i (25.0 mg,0.051 mmol) in CH₂Cl₂ (2.0 mL) was added 5520c (2.5 mL, 0.135 mmol),followed by DIPEA (68 uL, 0.4 mmol). The reaction mixture was stirred atroom temperature for 1.2 h, diluted with ethyl acetate (20.0 mL), washedwith 3% citric acid, brine, dried over NaHCO₃, filtered, concentratedand purified over SiO₂ using acetone-hexane (1:9) to yield 5665 (10.0mg). LCMS 646.2 (M+H).

A number of analogs of 5665 described in Table 2 were prepared using5520c according to the procedures described for the preparation of 5648.

Preparation of Compound 5688

Amine 5004a (998.0 mg, 4.6 mmol) was converted into the correspondingisocyanate 5030a using the procedure of step 5 in the preparation of5648.

To the isocyanate in CH₂Cl₂ (10.0 mL) was added N-methyl chloropropylamine 5030b (Aldrich, 490.0 mg, 4.6 mmol) and the reaction mixture wasstirred at room temperature for 12 h, washed with sat'd NaHCO₃, brine,dried over MgSO₄, filtered, concentrated and purified over SiO₂ usingethyl acetate/hexane to give 5530c (1.6 g, 4.6 mmol) in 100% yield.

Compound 5530c was converted to 5530d using the experimental proceduresStep 2 and Step 3 in the preparation of compound 5665.

To a cooled solution (0° C.) of the amine hydrochloride 5500i (25.0 mg,0.051 mmol) in CH₂Cl₂ (2.0 mL) was added 5530d (2.5 mL, 0.135 mmol),followed by DIPEA (68 uL, 0.4 mmol). The reaction mixture was stirred atroom temperature for 1.2 h, diluted with ethyl acetate (20.0 mL), washedwith 3% citric acid, brine, dried over NaHCO₃, filtered, concentratedand purified over SiO₂ using acetone-hexane (1:9) to yield 5688 (17.0mg). LCMS 660.2 (M+H).

A number of analogs of compound 5688 in Table 2 were prepared from 5530dusing the procedures described above.

Preparation of Compound 5700

Compound 3-chloro-propanol (181 uL, 2.51 mmol) was converted into thecorresponding chloroformate 5540b using the procedure step 5 in thepreparation of 5648.

To the ice-cooled chloroformate in THF (10.0 mL) was added amine 5004a(543.0 mg, 2.51 mmol) and the reaction mixture was stirred at roomtemperature for 12 h, diluted with EtOAc (250.0 mL) washed with sat'dNaHCO₃, brine, dried over MgSO₄, filtered, concentrated to yield 5540c(664 mg, 1.97 mmol). The crude was used directly in the next step.

Compound 5540c was converted to 5540d using the experimental proceduresfrom Step 2 and Step 3 in the preparation of compound 5665.

To a cooled solution (0° C.) of the amine hydrochloride 5500i (30.0 mg,0.0.6 mmol) in CH₂Cl₂ (2.0 mL) was added 5540d (2.6 mL, 0.131 mmol),followed by DIPEA (91 uL, 0.52 mmol). The reaction mixture was stirredat room temperature for 1.2 h, diluted with ethyl acetate (20.0 mL),washed with 3% citric acid, brine, dried over NaHCO₃, filtered,concentrated and purified over SiO₂ using acetone-hexane (1:9) to yield5700 (28.0 mg). LCMS 647.2 (M+H).

A number of analogs of 5700 in Table 2 were prepared from 5540d usingprocedures described above.

Preparation of Target Compound 5743

To a solution of 5003b (1.0 g, 2.9 mmol) in MeOH at −4° C. was addedNaBH₄ (11.52 mmol, 430.0 mg). After stirring for 20 min, the reactionwas quenched with CH₂Cl₂/sat'd NaHCO₃ (1:1, 60 mL). The aqueous layerwas extracted with DCM (3×20 mL). The organic layer was dried over MgSO₄and concentrated to yield a white solid which was used directly in thenext step. The crude from the previous step was re-dissolved in EtOH(50.0 mL) and Pd/C (10% by weight, 200 mg) was added. The reactionmixture was stirred under a H₂ atmosphere for 12 h, filtered over a padof celite and concentrated to yield 5550a (0.99 g).

Compound 5550a was converted to 5550b using the experimental proceduresfrom Step 3 in the preparation of compound 5665.

To a cooled solution (0° C.) of the amine hydrochloride 5550d (45.0 mg,0.094 mmol) in CH₂Cl₂ (2.0 mL) was added 5550b (2.8 mL, 2.0 mmol),followed by DIPEA (100 uL, 0.52 mmol). The reaction mixture was stirredat room temperature for 1.2 h, diluted with ethyl acetate (0.0 mL),washed with 3% citric acid, brine, dried over NaHCO₃, filtered,concentrated and purified over SiO₂ using EtOAc-CH₂Cl₂ (1:9 to 9:1) toyield product 5743 (32.0 mg). LCMS 705.2 (M+H).

A number of analogs of 5743 in Table 2 were prepared from 5550b usingprocedures described above.

Preparation of Compound 5754

The isocyanate 5754a was prepared from amine 5019 according to theprocedures described for the preparation of 5052d from 5052c.

The product 5754 was prepared from 5754a and 5052f according to theprocedures described for the preparation of compound 5237.

A number of analogs of 5754 in Table 2 were prepared from 5754a usingprocedures described above.

Preparation of Compound 5812

To the amine, (1.2 g, 5.5 mmol, 1 equiv.) in DCM (50 mL) was added 50 mlof sat. NaHCO₃. Stirred vigorously at ice temperature for 5 min. Stoppedstirring and phosgene (2 equiv., 11.09 mmol, 20% in toluene, 5.96 mL)was syringed out to the lower layer and restored the vigorous stirringimmediately. Separated the layers after 1 h. Washed the water layer onemore time with DCM (3 ml) and dried over sodium sulfate. Filtered andevaporated at high vac. with out hot bath to half the volume and thenpurged N₂ for 15 minutes. Diluted to 100 mL in DCM. Used as it is forfurther reaction.

To the amine 5812b (Aldrich, 0.5 g, 4.2 mmol, 1 equiv.) in methanol (20mL) was added the isocyanate (5.5 mmol, 1.3 equiv.) and triethyl amine(3.4 ml, 6 equiv, 25.2 mmol) and refluxed the reaction mixture at 90° C.for 48 h. Stirring was continued at 100° C. for another 5 h. The mixturewas concentrated and purified through a flash column chromatography togive product 5812c in 96.7% yield.

To compound 5812c (650 mg) was added 4 M HCl/Dioxane (25 mL) and stirredat room temperature for 0.5 h. Evaporated off the solvent and azeotropedwith hexane and then with ether. Kept under high vac for 4 h to give theproduct in quantitative yield.

To the amine hydrochloride (20 mg, 0.08 mmol, 1.3 equiv.) in DCM (5 ml)was added DIPEA (6 equiv.) at 0° C. Added isocyanate (3 mL, 0.02M inDCM) under N₂ atmosphere. After stirred for 30 min at 0° C. and 90 minat room temperature. Quenched with citric acid and extracted with EtOAcand washed with brine. Dried over anhydrous sodium sulfate and filteredand evaporated off the solvent. The crude product was purified via flashcolumn chromatography (10–40% acetone-hexane) to give compound 5812 in41% yield.

Representative compounds of the invention which exhibit excellent HCVprotease inhibitory activity are listed below in Tables 1 and 2 alongwith their biological activity in HCV continuous assay (ranges of Ki*values in nanomolar, nM): category A≦50 nM; category B>50 nM.

TABLE 1 Compound mol number structure Ki* weight 5106

A 652.86009 5107

A 678.89833 5108

B 720.85847 5109

A 638.833 5110

A 652.86009 5111

A 680.91427 5112

A 694.94136 5113

A 652.86009 5114

A 692.92542 5115

A 680.91427 5116

A 666.88718 5117

A 666.88718 5118

A 708.84732 5119

A 734.88556 5120

A 640.84894 5121

A 694.94136 5122

A 654.87603 5123

A 668.90312 5124

A 668.90312 5125

A 690.90948 5126

A 709.95603 5127

A 721.96718 5128

A 683.91779 5129

A 816.03767 5130

A 681.90185 5131

A 695.92894 5132

A 723.98312 5133

A 720.9796 5134

A 706.95251 5135

A 718.96366 5136

A 680.91427 5137

A 669.8907 5138

A 692.92542 5139

A 734.98463 5140

A 678.89833 5141

A 760.9238 5142

A 692.92542 5143

A 732.99075 5144

A 706.95251 5145

A 692.92542 5146

A 681.90185 5147

A 667.87476 5148

A 667.87476 5149

A 653.84767 5150

A 695.92894 5151

A 706.93045 5152

A 720.95754 5153

A 746.99578 5154

A 749.01172 5155

A 705.92415 5156

A 691.89706 5157

A 749.90023 5158

A 707.94009 5159

A 721.96718 5160

A 683.91779 5161

A 708.94639 5162

A 742.94233 5163

A 728.91524 5164

A 742.98596 5165

A 740.97002 5166

A 723.86199 5167

A 681.90185 5168

A 719.95124 5169

A 744.97984 5170

A 738.95408 5171

A 728.95887 5172

A 700.90469 5173

A 714.93178 5174

A 754.99711 5175

A 720.9796 5176

A 732.99075 5177

A 761.02287 5178

A 775.04996 5179

A 773.03402 5180

A 787.06111 5181

A 702.92063 5182

A 747.01784 5183

A 735.00669 5184

A 746.99578 5185

A 732.96869 5186

A 692.92542 5187

A 706.95251 5188

A 730.97481 5189

A 694.94136 5190

A 720.9796 5191

A 708.96845 5192

A 734.98463 5193

A 680.91427 5194

A 771.01808 5195

A 704.93657 5196

A 706.95251 5197

A 666.88718 5198

A 762.03251 5199

A 707.94009 5200

A 668.90312 5201

A 749.97773 5202

A 723.93949 5203

A 709.9124 5204

A 750.02136 5205

A 738.01021 5206

A 761.02287 5207

B 759.00693 5208

A 746.99578 5209

A 756.99099 5210

A 709.95603 5211

A 718.9416 5212

A 732.96869 5213

B 707.94009 5214

A 720.95754 5215

A 667.87476 5216

B 695.92894 5217

B 709.95603 5218

A 695.92894 5219

A 723.98312 5220

A 709.95603 5221

A 764.99232 5222

A 769.0242 5223

A 754.99711 5224

A 720.9796 5225

A 735.00669 5226

A 726.94293 5227

A 740.97002 5228

A 732.99075 5229

A 767.00826 5230

A 730.97481 5231

A 692.93 5232

A 706.95 5233

B 781.04 5234

B 747.02 5235

A 735.99 5236

A 708.97 5237

A 735.99 5238

A 748.01 5239

A 762.03 5240

A 750.02 5241

A 735.99 5242

A 745.99 5243

A 707.94009 5244

A 721.96718 5245

A 763.03881 5246

A 783.82184 5247

A 750.79191 5248

A 769.0242 5249

A 767.00826 5250

A 754.99711 5251

A 757.01305 5252

A 789.8476 5253

A 726.94293 5254

A 740.97002 5255

A 775.04996 5256

A 789.07705 5257

A 781.03535 5258

A 783.05129 5259

A 737.00057 5260

A 769.0242 5261

A 787.06111 5262

A 761.02287 5263

A 781.03535

TABLE 2 Compound number structure Ki* mol weight 5603

A 612.77596 5604

A 626.80305 5605

A 640.83014 5606

A 694.80143 5607

B 652.84129 5608

B 656.87317 5609

B 658.88911 5610

A 654.85723 5611

A 604.75306 5612

A 618.78015 5613

A 720.91717 5614

A 670.90026 5615

B 666.86838 5616

A 686.77853 5617

A 646.83433 5618

A 678.8359 5619

A 706.89008 5620

A 644.86202 5621

B 658.88911 5622

A 630.83493 5623

A 656.87317 5624

A 604.79669 5625

B 674.84488 5626

A 666.82475 5627

A 646.83433 5628

A 632.80724 5629

A 618.78015 5630

A 700.80562 5631

A 660.86142 5632

A 630.83493 5633

A 644.86202 5634

A 718.90123 5635

A 692.86299 5636

A 670.85663 5637

A 658.84548 5638

A 630.7913 5639

A 644.81839 5640

A 644.81839 5641

A 670.90026 5642

A 684.92735 5643

A 630.83493 5644

A 644.86202 5645

A 658.88911 5646

A 618.78015 5647

A 672.87257 5648

A 640.83014 5649

A 626.80305 5650

A 654.85723 5651

A 668.88432 5652

A 672.9162 5653

A 694.87893 5654

A 668.88432 5655

A 666.86838 5656

B 671.88784 5657

A 693.85057 5658

A 719.88881 5659

A 693.85057 5660

A 679.82348 5661

A 667.81233 5662

A 712.8604 5663

A 673.90378 5664

A 659.87669 5665

A 645.8496 5667

A 631.82251 5668

A 680.89547 5669

A 716.88529 5670

A 684.88372 5671

A 672.87257 5672

A 698.91081 5673

A 686.89966 5674

A 644.81839 5675

A 658.84548 5676

A 646.83433 5677

B 732.92832 5678

A 718.90123 5679

B 730.91238 5680

A 692.86299 5681

A 690.84705 5682

A 704.87414 5683

B 687.93087 5684

B 685.91493 5685

B 673.90378 5686

A 645.8496 5687

B 699.94202 5688

A 659.87669 5689

A 674.84488 5690

A 688.87197 5691

A 686.85603 5692

B 727.87507 5693

B 699.94202 5694

A 700.88312 5695

A 660.86142 5696

A 728.89644 5697

A 660.81779 5698

A 632.80724 5699

A 648.80664 5700

A 646.83433 5701

A 646.7907 5702

A 672.87257 5703

A 674.88851 5704

A 698.91081 5705

A 712.9379 5706

B 683.89899 5707

A 706.93371 5708

A 710.92196 5709

A 684.88372 5710

A 684.92735 5711

A 671.88784 5712

A 685.91493 5713

A 685.91493 5714

A 696.9385 5715

A 670.85663 5716

A 698.95444 5717

A 710.96559 5718

A 692.86299 5719

A 686.94329 5720

A 738.89864 5721

A 694.92256 5722

A 730.95601 5723

A 724.95 5724

A 718.94 5725

A 732.97 5726

A 729.93 5727

A 731.94 5728

A 733.96 5729

A 720.96 5730

A 698.95 5731

A 684.93 5732

A 696.94 5733

A 720.92 5734

A 660.91 5735

A 658.89 5736

A 710.97 5737

A 734.94 5738

B 746.96 5739

A 732.93 5740

A 694.92 5741

A 730.91 5742

B 692.91 5743

A 704.92 5744

B 718.94 5745

A 664.85 5746

B 694.92256 5747

A 746.87791 5748

A 706.93371 5749

A 702.90183 5750

A 690.89068 5751

A 718.94486 5752

A 727.73554 5753

A 714.95384 5754

A 712.9379 5755

A 726.96499 5756

A 755.78972 5757

A 700.92675 5758

A 702.94269 5759

A 710.92196 5760

A 761.75305 5761

A 672.87257 5762

A 686.89966 5763

A 772.91615 5764

A 728.94007 5765

A 732.97195 5766

A 718.94486 5767

A 720.9608 5768

A 730.95601 5769

A 744.9831 5770

A 723.87706 5771

A 737.90415 5772

A 735.88821 5773

A 695.82288 5774

A 709.84997 5775

A 738.97614 5776

A 726.96499 5777

A 740.99208 5778

A 714.95384 5779

A 724.94905 5780

A 708.94965 5781

A 712.98153 5782

A 710.96559 5783

A 698.95444 5784

A 712.9379 5785

A 736.9602 5786

A 698.91081 5787

A 728.94007 5788

A 753.00323 5789

A 718.94486 5790

A 700.97038 5791

A 720.9608 5792

A 698.95444 5793

A 779.04147 5794

A 767.03032 5795

A 781.05741 5796

A 738.97614 5797

A 765.01438 5798

A 726.96499 5799

A 712.9379 5800

A 724.94905 5801

A 740.99208 5802

A 755.01917 5803

A 753.00323 5804

A 669.82827 5805

A 673.86015 5806

A 700.92675 5807

A 716.96978 5808

A 743.00802 5809

A 740.99208 5810

A 767.03032 5811

A 631.77888 5812

A 683.85536 5813

A 687.88724 5814

A 671.84421 5815

A 685.8713 5816

A 685.8713 5817

A 740.99208 5818

A 753.00323

The present invention relates to novel HCV protease inhibitors. Thisutility can be manifested in their ability to inhibit the HCV NS3/NS4aserine protease. A general procedure for such demonstration isillustrated by the following in vitro assay.

Assay for HCV Protease Inhibitory Activity:

Spectrophotometric Assay:

Spectrophotometric assay for the HCV serine protease can be performed onthe inventive compounds by following the procedure described by R. Zhanget al, Analytical Biochemistry, 270 (1999) 268–275, the disclosure ofwhich is incorporated herein by reference. The assay based on theproteolysis of chromogenic ester substrates is suitable for thecontinuous monitoring of HCV NS3 protease activity. The substrates arederived from the P side of the NS5A-NS5B junction sequence(Ac-DTEDVVX(Nva), where X=A or P) whose C-terminal carboxyl groups areesterified with one of four different chromophoric alcohols (3- or4-nitrophenol, 7-hydroxy-4-methyl-coumarin, or 4-phenylazophenol).Illustrated below are the synthesis, characterization and application ofthese novel spectrophotometric ester substrates to high throughputscreening and detailed kinetic evaluation of HCV NS3 proteaseinhibitors.

Materials and Methods:

Materials: Chemical reagents for assay related buffers are obtained fromSigma Chemical Company (St. Louis, Mo.). Reagents for peptide synthesiswere from Aldrich Chemicals, Novabiochem (San Diego, Calif.), AppliedBiosystems (Foster City, Calif.) and Perseptive Biosystems (Framingham,Mass.). Peptides are synthesized manually or on an automated ABI model431A synthesizer (from Applied Biosystems). UV/VIS Spectrometer modelLAMBDA 12 was from Perkin Elmer (Norwalk, Conn.) and 96-well UV plateswere obtained from Corning (Corning, N.Y.). The prewarming block can befrom USA Scientific (Ocala, Fla.) and the 96-well plate vortexer is fromLabline Instruments (Melrose Park, Ill.). A Spectramax Plus microtiterplate reader with monochrometer is obtained from Molecular Devices(Sunnyvale, Calif.).

Enzyme Preparation:

Recombinant heterodimeric HCV NS3/NS4A protease (strain 1a) is preparedby using the procedures published previously (D. L. Sali et al,Biochemistry, 37 (1998) 3392–3401). Protein concentrations aredetermined by the Biorad dye method using recombinant HCV proteasestandards previously quantified by amino acid analysis. Prior to assayinitiation, the enzyme storage buffer (50 mM sodium phosphate pH 8.0,300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside and 10 mM DTT) isexchanged for the assay buffer (25 mM MOPS pH 6.5, 300 mM NaCl, 10%glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5 μM DTT) utilizing aBiorad Bio-Spin P-6 prepacked column.

Substrate Synthesis and Purification:

The synthesis of the substrates is done as reported by R. Zhang et al,(ibid.) and is initiated by anchoring Fmoc-Nva-OH to 2-chlorotritylchloride resin using a standard protocol (K. Barlos et al, Int. J. Pept.Protein Res., 37 (1991), 513–520). The peptides are subsequentlyassembled, using Fmoc chemistry, either manually or on an automatic ABImodel 431 peptide synthesizer. The N-acetylated and fully protectedpeptide fragments are cleaved from the resin either by 10% acetic acid(HOAc) and 10% trifluoroethanol (TFE) in dichloromethane (DCM) for 30min, or by 2% trifluoroacetic acid (TFA) in DCM for 10 min. The combinedfiltrate and DCM wash is evaporated azeotropically (or repeatedlyextracted by aqueous Na₂CO₃ solution) to remove the acid used incleavage. The DCM phase is dried over Na₂SO₄ and evaporated.

The ester substrates are assembled using standard acid-alcohol couplingprocedures (K. Holmber et al, Acta Chem. Scand., B33 (1979) 410–412).Peptide fragments are dissolved in anhydrous pyridine (30–60 mg/ml) towhich 10 molar equivalents of chromophore and a catalytic amount (0.1eq.) of para-toluenesulfonic acid (pTSA) were added.Dicyclohexylcarbodiimide (DCC, 3 eq.) is added to initiate the couplingreactions. Product formation is monitored by HPLC and can be found to becomplete following 12–72 hour reaction at room temperature. Pyridinesolvent is evaporated under vacuum and further removed by azeotropicevaporation with toluene. The peptide ester is deprotected with 95% TFAin DCM for two hours and extracted three times with anhydrous ethylether to remove excess chromophore. The deprotected substrate ispurified by reversed phase HPLC on a C3 or C8 column with a 30% to 60%acetonitrile gradient (using six column volumes). The overall yieldfollowing HPLC purification can be approximately 20–30%. The molecularmass can be confirmed by electrospray ionization mass spectroscopy. Thesubstrates are stored in dry powder form under desiccation.

Spectra of Substrates and Products:

Spectra of substrates and the corresponding chromophore products areobtained in the pH 6.5 assay buffer. Extinction coefficients aredetermined at the optimal off-peak wavelength in 1-cm cuvettes (340 nmfor 3-Np and HMC, 370 nm for PAP and 400 nm for 4-Np) using multipledilutions. The optimal off-peak wavelength is defined as that wavelengthyielding the maximum fractional difference in absorbance betweensubstrate and product (product OD−substrate OD)/substrate OD).

Protease Assay:

HCV protease assays are performed at 30° C. using a 200 μl reaction mixin a 96-well microtiter plate. Assay buffer conditions (25 mM MOPS pH6.5, 300 mM NaCl, 10% glycerol, 0.05% lauryl maltoside, 5 μM EDTA and 5μM DTT) are optimized for the NS3/NS4A heterodimer (D. L. Sali et al,ibid.)). Typically, 150 μl mixtures of buffer, substrate and inhibitorare placed in wells (final concentration of DMSO≦4% v/v) and allowed topreincubate at 30° C. for approximately 3 minutes. Fifty μls ofprewarmed protease (12 nM, 30° C.) in assay buffer, is then used toinitiate the reaction (final volume 200 μl). The plates are monitoredover the length of the assay (60 minutes) for change in absorbance atthe appropriate wavelength (340 nm for 3-Np and HMC, 370 nm for PAP, and400 nm for 4-Np) using a Spectromax Plus microtiter plate readerequipped with a monochrometer (acceptable results can be obtained withplate readers that utilize cutoff filters). Proteolytic cleavage of theester linkage between the Nva and the chromophore is monitored at theappropriate wavelength against a no enzyme blank as a control fornon-enzymatic hydrolysis. The evaluation of substrate kinetic parametersis performed over a 30-fold substrate concentration range (˜6–200 μM).Initial velocities are determined using linear regression and kineticconstants are obtained by fitting the data to the Michaelis-Mentenequation using non-linear regression analysis (Mac Curve Fit 1.1, K.Raner). Turnover numbers (k_(cat)) are calculated assuming the enzyme isfully active.

Evaluation of Inhibitors and Inactivators:

The inhibition constants (K_(i)) for the competitive inhibitorsAc-D-(D-Gla)-L-1-(Cha)-C—OH (27), Ac-DTEDVVA(Nva)-OH andAc-DTEDVVP(Nva)-OH are determined experimentally at fixed concentrationsof enzyme and substrate by plotting v_(o)/v_(i) vs. inhibitorconcentration ([I]_(o)) according to the rearranged Michaelis-Mentenequation for competitive inhibition kinetics:v_(o)/v_(i)=1+[I]_(o)/(K_(i)(1+[S]_(o)/K_(m))), where v_(o) is theuninhibited initial velocity, v_(i) is the initial velocity in thepresence of inhibitor at any given inhibitor concentration ([I]_(o)) and[S]_(o) is the substrate concentration used. The resulting data arefitted using linear regression and the resulting slope,1/(K_(i)(1+[S]_(o)/K_(m)), is used to calculate the K_(i) value. The Ki*values (in nanoMolar) for some of the inventive compounds are in thefollowing Table 4:

TABLE 4 Compound number structure Ki* 5619

11 5652

19 5705

3 5724

19 5731

15 5753

4 5754

4 5755

7 5775

5 5781

9 5793

11 5796

3.2

While the present invention has been described with in conjunction withthe specific embodiments set forth above, many alternatives,modifications and other variations thereof will be apparent to those ofordinary skill in the art. All such alternatives, modifications andvariations are intended to fall within the spirit and scope of thepresent invention.

1. A compound, or enantiomers, stereoisomers, rotamers, tautomers, andracemates of said compound, or a pharmaceutically acceptable salt,solvate or ester of said compound, said compound having the generalstructure shown in Formula I:

wherein: R¹ is H, OR⁸, NR⁹R¹⁰, or CHR⁹R¹⁰, wherein R⁸, R⁹ and R¹⁰ can bethe same or different, each being independently selected from the groupconsisting of H, alkyl-, alkenyl-, alkynyl-, aryl-, heteroalkyl-,heteroaryl-, cycloalkyl-, heterocyclyl-, arylalkyl-, andheteroarylalkyl; A and M are connected to each other (in other words,A-E-L-M taken together) such that the moiety:

shown above in Formula I forms either a three, four, six, seven oreight-membered cycloalkyl, E is C(H) or C(R); L is C(H) or C(R); R, R²,and R³ can be the same or different, each being independently selectedfrom the group consisting of H, alkyl-, alkenyl-, alkynyl-, cycloalkyl-,heteroalkyl-, heterocyclyl-, aryl-, heteroaryl-, (cycloalkyl)alkyl-,(heterocyclyl)alkyl-, aryl-alkyl-, and heteroaryl-alkyl-; and Y isselected from the following moieties:

 wherein G is NH or O, and R¹⁵, R¹⁷, R¹⁸, R¹⁹ and R²⁰ can be the same ordifferent, each being independently selected from the group consistingof H, C₁–C₁₀ alkyl, C₁–C₁₀ heteroalkyl, C₂–C₁₀ alkenyl, C₂–C₁₀heteroalkenyl, C₂–C₁₀ alkynyl, C₂–C₁₀ heteroalkynyl, C₃–C₈ cycloalkyl,C₃–C₈ heterocyclyl, aryl, heteroaryl, or alternately: (i) R¹⁵ and R¹⁹are connected to each other to form a five to eight-membered cycloalkylor heterocyclyl, or R¹⁵ and R²⁰ are connected to each other to form afive to eight-membered cycloalkyl or heterocyclyl, and (ii) likewise,independently, R¹⁷ and R¹⁸ are connected to each other to form a threeto eight-membered cycloalkyl or heterocyclyl, wherein each of saidalkyl, aryl, heteroaryl, cycloalkyl or heterocyclyl can be unsubstitutedor optionally independently substituted with one or more moietiesselected from the group consisting of: hydroxy, alkoxy, aryloxy, thio,alkylthio, arylthio, amino, amido, alkylamino, arylamino, alkylsulfonyl,arylsulfonyl, sulfonamido, alkylsulfonamido, arylsulfonamido, keto,carboxy, carbalkoxy, carboxamido, alkoxycarbonylamino,alkoxycarbonyloxy, alkylureido, arylureido, halo, cyano, and nitro andwherein any carbon or heteroatom with unsatisfied valances in the text,schemes, examples and tables herein is assumed to have hydrogen atom(s)to satisfy the valences.
 2. The compound of claim 1, wherein R¹ isNR⁹R¹⁰, and R⁹ is H, R¹⁰ is H, alkyl, aryl, heteroalkyl, heteroaryl,cycloalkyl, alkyl-aryl, alkyl-heteroaryl, aryl-alkyl, alkenyl, alkynylor heteroaryl-alkyl.
 3. The compound of claim 2, wherein R¹⁰ is selectedfrom the group consisting of:


4. The compound of claim 1, wherein R² is selected from the groupconsisting of the following moieties:


5. The compound of claim 1, wherein R³ is selected from the groupconsisting of:

wherein R³¹ is OH or O-alkyl; and R³² is H, C(O)CH₃, C(O)OtBu orC(O)N(H)tBu.
 6. The compound of claim 5, wherein R³ is selected from thegroup consisting of the following moieties:


7. The compound of claim 1, wherein G is NH.
 8. The compound of claim 1,wherein Y is selected from the group consisting of:

wherein Y³⁰ and Y³¹ are selected from the group consisting of:

Y³² is selected from the group consisting of:

and Y¹² is selected from H, COOH, COOMe, CONH₂, OMe, OH, OCF₃,OCH(CH₃)₂, OC(CH₃)₃, F, Cl, Br, NH₂, NHSO₂CH₃, NHC(O)CH₃, NHCO₂CH₃, NO₂,SO₂NH₂, CF₃, Me, Et, isopropyl, cyclopropyl, t-butyl and phenyl.
 9. Thecompound of claim 8, wherein Y is selected from the group consisting of:

wherein Y³⁰ and Y³¹ are selected from the group consisting of:

Y³² is selected from the group consisting of:

and Y¹² is selected from H, COOH, COOMe, CONH₂, OMe, OH, OCF₃,OCH(CH₃)₂, OC(CH₃)₃, F, Cl, Br, NH₂, NHSO₂CH₃, NHC(O)CH₃, NHCO₂CH₃, NO₂,SO₂NH₂, CF₃, Me, Et, isopropyl, cyclopropyl, t-butyl and phenyl.
 10. Thecompound of claim 1, wherein the moiety:

is selected from the following structures:


11. The compound of claim 10, wherein the moiety:

is selected from the following structures:


12. The compound of claim 11, wherein the moiety:

is selected from the following structures:


13. The compound of claim 1, wherein R¹ is NHR¹⁰, where R¹⁰ is selectedfrom the group consisting of:

R² is selected from the group consisting of the following moieties:

R³ is selected from the group consisting of the following moieties:

Y is selected from the group consisting of:

 wherein Y³⁰ and Y³¹ can be the same or different, each beingindependently selected from the group consisting of:

 wherein Y³² is selected from the group consisting of:

and Y¹² is selected from H, COOH, COOMe, CONH₂, OMe, OH, OCF₃,OCH(CH₃)₂, OC(CH₃)₃, F, Cl, Br, NH₂, NHSO₂CH₃, NHC(O)CH₃, NHCO₂CH₃, NO₂,SO₂NH₂, CF₃, Me, Et, isopropyl, cyclopropyl, t-butyl, or phenyl; and themoiety:


14. A pharmaceutical composition comprising as an active ingredient atleast one compound of claim
 1. 15. The pharmaceutical composition ofclaim 14 additionally comprising at least one pharmaceuticallyacceptable carrier.
 16. The pharmaceutical composition of claim 15,additionally containing at least one antiviral agent.
 17. Thepharmaceutical composition of claim 16, still additionally containing atleast one interferon.
 18. The pharmaceutical composition of claim 17,wherein said at least one antiviral agent is ribavirin and said at leastone interferon is α-interferon or pegylated interferon.
 19. A method oftreating Hepatitis C Virus (“HCV”), said method comprising administeringto a patient in need of such treatment a pharmaceutical compositionwhich comprises therapeutically effective amounts of at least onecompound of claim
 1. 20. The method of claim 19, wherein saidadministration is oral or subcutaneous.
 21. A method of preparing apharmaceutical composition for treating Hepatitis C Virus (“HCV”), saidmethod comprising bringing into intimate physical contact at least onecompound of claim 1 and at least one pharmaceutically acceptablecarrier.
 22. A compound of claim 1 exhibiting Hepatitis C Virus (“HCV”)protease inhibitory activity, or enantiomers, stereoisomers, rotamers,tautomers, and racemates of said compound, or a pharmaceuticallyacceptable salt, solvate or ester of said compound, said compound beingselected from the compounds of structures listed below:


23. A pharmaceutical composition hepatitis C virus (“HCV”), saidcomposition comprising therapeutically effective amount of one or morecompounds in claim 22 and a pharmaceutically acceptable carrier.
 24. Thepharmaceutical composition of claim 23, additionally containing at leastone antiviral agent.
 25. The pharmaceutical composition of claim 24,additionally containing at least one interferon or PEG-interferon alphaconjugate (“pegylated interferon”).
 26. The pharmaceutical compositionof claim 25, wherein said at least one antiviral agent is ribavirin andsaid at least one interferon is α-interferon or pegylated interferon.27. A method of treatment of a hepatitis C virus, comprisingadministering an effective amount of one or more compounds of claim 22.28. A method of treating HCV, said method comprising administering to apatient in need of such treatment, a pharmaceutical composition whichcomprises therapeutically effective amounts of at least one compound, orenantiomers, stereoisomers, rotamers, tautomers, and racemates of saidcompound, or a pharmaceutically acceptable salt, solvate or ester ofsaid compound, said compound being selected from the following:


29. A compound of claim 1 in purified form.